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PubMed2003

Design of efficient, broadband single-element (20-80 MHz) ultrasonic transducers for medical imaging applications.

This paper discusses the design, fabrication, and testing of sensitive broadband lithium niobate (LiNbO3) single-element ultrasonic transducers in the 20-80 MHz frequency range. Transducers of varying dimensions were built for an f# range of 2.0-3.1. The desired focal depths were achieved by either casting an acoustic lens on the transducer face or press-focusing the piezoelectric into a spherical curvature. For designs that required electrical impedance matching, a low impedance transmission line coaxial cable was used. All transducers were tested in a pulse-echo arrangement, whereby the center frequency, bandwidth, insertion loss, and focal depth were measured. Several transducers were fabricated with center frequencies in the 20-80 MHz range with the measured -6 dB bandwidths and two-way insertion loss values ranging from 57 to 74% and 9.6 to 21.3 dB, respectively. Both transducer focusing techniques proved successful in producing highly sensitive, high-frequency, single-element, ultrasonic-imaging transducers. In vivo and in vitro ultrasonic backscatter microscope (UBM) images of human eyes were obtained with the 50 MHz transducers. The high sensitivity of these devices could possibly allow for an increase in depth of penetration, higher image signal-to-noise ratio (SNR), and improved image contrast at high frequencies when compared to previously reported results.

Computer-Aided Design↗
PubMed2005

The energy method for analyzing the piezoelectric electroacoustic transducers.

The energy method of calculating the properties of piezoelectric electroacoustic transducers is considered. The Lagrangian of an electroacoustic transducer as a system performing multiple energy conversions is introduced. The Euler equations describing operation of a transducer with many mechanical degrees of freedom are derived from the least action principle. The corresponding multicontour equivalent circuit of the transducer is introduced. For the particular case of a transducer having one mechanical degree of freedom the governing equations are obtained by applying the Energy Conservation Law, and equivalent circuit with one mechanical branch also introduced. Application of the energy method is illustrated with examples of the pulsating spherical transducer as the one degree of freedom system and the multimode cylindrical transducer comprised of circular rings as the system with multiple degrees of freedom. Advantages of the method for application with electroacoustic transducers are summarized.

Acoustics↗
PubMed1992

Electromagnetic midsagittal articulometer systems for transducing speech articulatory movements.

This paper describes two electromagnetic midsagittal articulometer (EMMA) systems that were developed for transducing articulatory movements during speech production. Alternating magnetic fields are generated by transmitter coils that are mounted in an assembly that fits on the head of a speaker. The fields induce alternating voltages in a number of small transducer coils that are attached to articulators in the midline plane, inside and outside the vocal tract. The transducers are connected by fine lead wires to receiver electronics whose output voltages are processed to yield measures of transducer locations as a function of time. Measurement error can arise with this method, because as the articulators move and change shape, the transducers can undergo a varying amount of rotational misalignment with respect to the transmitter axes; both systems are designed to correct for transducer misalignment. For this purpose, one system uses two transmitters and biaxial transducers; the other uses three transmitters and single-axis transducers. The systems have been compared with one another in terms of their performance, human subjects compatibility, and ease of use. Both systems can produce useful midsagittal-plane data on articular movement, and each one has a specific set of advantages and limitations. (Two commercially available systems are also described briefly for comparison purposes). If appropriate experimental controls are used, the three-transmitter system is preferable for practical reasons.

Biomechanical Phenomena↗
PubMed1990

Percutaneous vs. transcutaneous transducers for hearing by direct bone conduction.

There is a substantial need for improvement of the hearing situation for patients, with chronic middle ear or ear canal disorders. To improve hearing for these patients, two different bone conduction hearing systems have been developed. The Nobelpharma Auditory System HC 200-the bone-anchored hearing aid we present here-uses a percutaneous transducer; whereas the Audiant device, developed by Dr. Jack Hough, uses a transcutaneous transducer. In percutaneous transmission, the transducer is directly coupled to the bone by means of a permanent skin penetration, whereas in transcutaneous transmission one part of the transducer is implanted and the other part is kept outside the intact skin and soft tissue. Comprehensive audiologic assessments indicate great differences in performance between the two systems. These differences probably originate in differences in length of gap and in different suspension properties of the two transducer systems. This article will demonstrate that large gaps, such as in the transcutaneous transducer, can be devastating for power consumption, maximum output capability, and second harmonic distortion. Since the properties of the suspension in the transcutaneous transducer are not under adequate control and the complication risk of permanent skin penetration is low, we continue to concentrate our efforts on percutaneous transducer systems.

Bone Conduction↗
PubMed2001

Invasive arterial BP monitoring in trauma and critical care: effect of variable transducer level, catheter access, and patient position.

OBJECTIVES: (1) To determine the validity of current recommendations for direct arterial BP measurement that suggest that the transducer (zeroed to atmosphere) be placed level with the catheter access regardless of subject positioning: and (2) to investigate the effect of transducer level, catheter access site, and subject positioning on direct arterial BP measurement. DESIGN: Prospective, controlled laboratory study. SETTING: Large animal laboratory. SUBJECTS: Five Yorkshire pigs. INTERVENTIONS: Anesthetized animals had 16F catheters placed at three access sites: aortic root, femoral artery, and distal hind limb. Animals were placed in supine, reverse Trendelenburg 35 degrees, and Trendelenburg 25 degrees positions with a transducer placed level to each access site while in every position. MEASUREMENTS AND MAIN RESULTS: For each transducer level, five systolic and diastolic pressures were measured and used to calculate five corresponding mean arterial pressures (MAPs) at each access site. When transducers were at the aortic root, MAP corresponding to aortic root pressure was obtained in all positions regardless of catheter access site. When transducers were moved to the level of catheter access, as current recommendations suggest, significant errors in aortic MAP occurred in the reverse Trendelenburg position. The same trend for error was noted in the Trendelenburg position but did not reach statistical significance. CONCLUSIONS: (1) Current recommendations that suggest placing the transducer at the level of catheter access regardless of patient position are invalid. Significant errors occur when subjects are in nonsupine positions. (2) Valid determination of direct arterial BP is dependent only on transducer placement at the level of the aortic root, and independent of catheter access site and patient position.

Animals↗
PubMed1991

Detection of renal stones with real-time sonography: effect of transducers and scanning parameters.

Three experiments with a variety of transducers and scanning parameters were designed to investigate if renal stones could be detected with greater certainty by using particular transducers or scanning parameters. First, the lateral resolution, derived from the -6-dB size of the beam profile, was measured at various depths for five transducers commonly used for renal sonography. Second, an in vitro test object was constructed from bovine liver, porcine kidneys, and two renal calculi to access gray-scale map effects on shadow visibility before and after storage in the digital scan converter. The third experiment combined 15 lithotripsy patients with known renal stones with 16 patients in whom the results of renal sonography and other radiographic procedures suggested renal calculi. The group of 15 patients was scanned several times with the transducers and gray-scale maps studied earlier, and the group of 16 patients was scanned only with one transducer and one gray-scale map. On radiographs, 12 of the 16 patients did not have renal calculi. Sonograms of the test object showed that low-contrast images were best for detection of posterior shadows. Three radiologists interpreted the 31 sonograms with a sensitivity of 81% and a specificity of 86% for detecting renal stones. For the 15 cases of renal stones scanned with a variety of transducers, the three radiologists found that annular-array transducers depicted stone shadowing with less ambiguity than mechanical sector transducers did 81% of the time.

Animals↗
PubMed2006

Effect of transducer velocity on intramuscular temperature during a 1-MHz ultrasound treatment.

STUDY DESIGN: A 3 x 2 repeated-measures design was used. The independent variables were transducer velocity (2-3 cm/s, 4-5 cm/s, and 7-8 cm/s) and time (pretreatment and posttreatment). OBJECTIVE: To determine if transducer velocity of a 1-MHz ultrasound treatment affects intramuscular tissue temperature. BACKGROUND: Most authors advocate ultrasound transducer velocities of 2 to 4 cm/s within an area of 2 to 3 times the effective radiating area or 2 times the size of the transducer head. However, a much faster rate of application (approximately 7-8 cm/s) is often observed in clinical settings. METHODS AND MEASURES: Eleven healthy screened volunteers (9 males, 2 females; mean +/- SD age, 22.6 +/- 1.7 years; mean +/- SD height, 175.7 +/- 13.7 cm; mean +/- SD body mass, 82.5 +/- 19.5 kg) were randomly assigned to a treatment order with all conditions administered during a single testing session. Each transducer velocity condition was administered for 10 minutes, using 1-MHz ultrasound with a 100% continuous duty cycle at an intensity of 1.5 W/cm2 over an area twice the size of the transducer head. After the first treatment, the 2 remaining subsequent velocity conditions were administered after the intramuscular temperature returned to within +/- 0.3 degrees C of the initial pretreatment temperature for 5 minutes. The dependent variable was left triceps surae muscle temperature measured at 3 cm below one half the measured skinfold thickness. RESULTS: Temperature increase across the 3 velocities was within 0.4 degrees C (F2.20 = 0.07, P = .93). Posttreatment values (mean +/- SD) ranged from 42.7 degrees C +/- 2.3 degrees C for the slowest velocity to 43.1 degrees C +/- 1.4 degrees C for the fastest velocity. Temperature increase was significant for time (F1.01 = 155.68, P<.00001), increasing from 37.8 degrees C +/- 0.8 degrees C pretreatment to 42.9 degrees C +/- 1.9 degrees C after treatment. CONCLUSION: Very similar intramuscular temperature increases can be observed among ultrasound treatments (10-minute duration, 1-MHz frequency, 100% continuous duty cycle, 1.5 W/cm2 intensity, within an area twice the size of the transducer head), with transducer velocities of 2 to 3, 4 to 5, and 7 to 8 cm/s.

Adult↗
PubMedDate not supplied

[Echographic study with high-frequency and high-spatial resolution transducer in the evaluation of renal transplant in pediatric age].

PURPOSE: We investigated the role of power Doppler US with a high-frequency and high-resolution transducer (13 MHz) in the visualization of interlobular arterioles in patients with normally functioning renal transplants or with chronic rejection. MATERIAL AND METHODS: We examined 15 patients (mean age 15 years; range 10-18 years) with a General Electric 500 MD unit using 7.5 and 13 MHz linear transducers. In all the patients serum creatinine and diuresis were evaluated; 4 patients underwent US-guided biopsy that resulted in the diagnosis of chronic rejection. RESULTS: Normally functioning renal transplants were found in 11 patients and chronic rejection was seen in 4. In normally functioning renal transplants, interlobular vessels could be depicted as "cortical blush" with the 7.5 MHz transducer; in the same patients power Doppler US with the 13 MHz transducer permitted a correct evaluation of interlobular vessels that were arranged in series like a palisade. In chronic rejection power Doppler US with the 13 MHz transducer better depicted cortical vascularity and showed irregular, narrow arteries. CONCLUSION: Power Doppler US with a 13 MHz transducer is particularly useful in children after renal transplants due to their reduced tissutal thickness. The lateral resolution of 13 MHz transducers (< 0.3 mm) allows to separate interlobular vessels from each other and the high frequency of the probe can depict interlobular vessels in the peripheral cortex. The optimal visualization of cortical vascularity with a 13 MHz transducer allows early detection of chronic rejection.

Adolescent↗
PubMed1986

The production of generalized transducing phage by bacteriophage lambda.

Generalized transduction has for about 30 years been a major tool in the genetic manipulation of bacterial chromosomes. However, throughout that time little progress has been made in understanding how generalized transducing particles are produced. The experiments presented in this paper use phage lambda to assess some of the factors that affect that process. The results of those experiments indicate: the production of generalized transducing particles by bacteriophage lambda is inhibited by the phage lambda exonuclease (Exo). Also inhibited by lambda Exo is the production of lambda docR particles, a class of particles whose packaging is initiated in bacterial DNA and terminated at the normal phage packaging site, cos. In contrast, the production of lambda docL particles, a class of particles whose packaging is initiated at cos and terminated in bacterial DNA, is unaffected by lambda Exo; lambda-generalized transducing particles are not detected in induced lysis-defective (S-) lambda lysogens until about 60-90 min after prophage induction. Since wild-type lambda would normally lyse cells by 60 min, the production of lambda-generalized transducing particles depends on the phage being lysis-defective; if transducing lysates are prepared by phage infection then the frequency of generalized transduction for different bacterial markers varies over a 10-20-fold range. In contrast, if transducing lysates are prepared by the induction of a lambda lysogen containing an excision-defective prophage, then the variation in transduction frequency is much greater, and markers adjacent to, and on both sides of, the prophage are transduced with much higher frequencies than are other markers; if the prophage is replication-defective then the increased transduction of prophage-proximal markers is eliminated; measurements of total DNA in induced lysogens indicate that part of the increase in transduction frequency following prophage induction can be accounted for by an increase in the amount of prophage-proximal bacterial DNA in the cell. Measurements of DNA in transducing particles indicate that the rest of the increase is probably due to the preferential packaging of the prophage-proximal bacterial DNA. These results are most easily interpreted in terms of a model for the initiation of bacterial DNA packaging by lambda, in which the proteins involved (Ter) do not recognize any particular sequence in bacterial DNA but rather recognize some feature of the DNA tht is sensitive to lambda exonuclease, such as a nick or a double-stranded cut.(ABSTRACT TRUNCATED AT 400 WORDS)

Bacteriophage lambda↗
PubMed2006

Decreased homing of retrovirally transduced human bone marrow CD34+ cells in the NOD/SCID mouse model.

OBJECTIVE: Many clinical gene therapy trials have described poor engraftment of retrovirally transduced CD34(+) cells. Because engraftment is dependent upon successful homing of graft cells to the bone marrow (BM), we examined whether retroviral-mediated gene transfer (RMGT) induces a homing defect in CD34(+) cells. METHODS: Homing of fluorescently labeled human BM CD34(+) cells transduced with three separate retroviral vectors (MFG-eGFP, LNC-eGFP, and LXSN) was assessed in nonobese diabetic/severe combined immunodeficient mice. RESULTS: Homing of transduced CD34(+) cells was significantly decreased 20 hours after transplantation compared with freshly isolated control and cultured untransduced control cells. Specifically, homing of GFP(+) cells in the graft was preferentially decreased thus skewing the contribution of transduced cells to engraftment. Transduced cells were not selectively trapped in other organs and BM-homed transduced cells did not undergo apoptosis at a higher rate than untransduced cells. Adhesion molecule expression and binding activity was not altered by RMGT. This homing defect was reversed when transduced cells were cultured over CH-296 for 2 additional days with SCF only. CONCLUSION: These data suggest that RMGT of hematopoietic cells may compromise their homing potential and implicate transduction-induced reduced homing in the observed low engraftment of retrovirally transduced CD34(+) cells. These results may have a direct clinical application in gene therapy protocols.

Animals↗
PubMed2004

Thermo-mechanical stress effect on 1-3 piezocomposite power transducer performance.

This paper deals with the emission performance of 1-3 piezoelectric composite power transducers made with a hard PZT (Navy III) and epoxy resins with a high glass-rubber transition temperature. Following the "dice and fill" technique, various composite transducers with 30 and 50% PZT volume fractions were fabricated with an air backing and no front matching layer with resonance operating frequencies around 500 kHz. The transducers were first evaluated under isothermal conditions, with a low emission duty cycle. Efficiencies as high as 95% were monitored as a function of the instantaneous input power up to a 60 W/cm(2) density. The effect of the polymer matrix mechanical losses and the fabrication conditions is then discussed. For the transducer thermal stability, the case of long duty cycle or continuous emission was considered in a second evaluation. In this case the transducer working temperature and axial radiated pressure were monitored as functions of the input power density up to 40 W/cm(2). It is shown that the transducer efficiency and working temperature were strongly dependent on the type of resin used but also on the PZT material, even for hard PZT compositions. A composite transducer configuration with strongly improved thermal stability was investigated demonstrating a working temperature higher than 90 degrees C and an extended power range (30-40 W/cm(2)). The composite thermal breakdown mechanism was analyzed and the effect of the curing-induced thermo-mechanical stresses on the PZT mechanical losses was considered in relation to the composite working temperature. Measurements of the composite mechanical losses versus the temperature were obtained and related to the variation of the PZT mechanical losses with the stresses due to the composite transducer temperature change. It is found that the thermally induced stress can strongly influence the PZT ceramic mechanical losses and that it can be the reason for a thermal breakdown taking place at a temperature much lower than the epoxy resin transition.

Journal Article↗
PubMed2006

Rare earth ultrasonic transducer technique research.

Transducer is the necessary and most important part in the maxonics. New applications demand for new transducer, and the renovation of the transducer will open a new field for the application of the maxonics. The important method to develop new transducer is to find and develop new material. The rare earth giant magnetostrictive material Terfenol-D is a new functional material with good performance. In this paper, the rare earth ultrasonic transducer was analyzed theoretically and devised in four-port method and FEM software ANSYS. A rare earth ultrasonic transducer and the transducer with the half wave horn are developed, whose performance has shown unique advantage compared with the piezoelectric transducer.

Journal Article↗
PubMed2004

In vivo selection for human and murine hematopoietic cells transduced with a therapeutic MGMT lentiviral vector that inhibits HIV replication.

We have developed an HIV-based lentiviral vector, VRX496, which efficiently transduces human CD34+ progenitors and CD4+ T lymphocytes. VRX496 contains an antisense sequence against the HIV envelope and is currently being evaluated for safety in a clinical trial for treatment of HIV. Selective outgrowth of transduced hematopoietic cells in vivo is anticipated to increase the therapeutic efficacy of this treatment by maximizing the persistence of virus-resistant cells in the body. Although HIV resistance is selective, additional selection may aid in treatment efficacy due to the vast quantity of target cells. Therefore, we engineered VRX496 to express the P140K MGMT gene to drive potent drug-mediated in vivo selection for transduced hematopoietic long-term repopulating cells. Suboptimally transduced T cell cultures treated with O6-benzylguanine and BCNU were selected from 3 to 100%, and after selection cultures did not support HIV replication. Primary CD34+ progenitors derived from G-CSF-mobilized peripheral blood were transduced at 27 to 35% efficiency. Approximate sixfold selection was observed for transduced CD34+ progenitors, colony-forming units, and long-term culture-initiating cells. Multilineage in vivo selection was demonstrated for transduced murine hematopoietic cells in human CD34(+)-derived hematopoietic cells in NOD-SCID mice. These results establish efficient ex vivo and in vivo selection for hematopoietic cells transduced with lentiviral vectors and support the potential therapeutic benefit of this strategy in human gene therapy.

Animals↗
PubMed2001

The superfamily of chemotaxis transducers: from physiology to genomics and back.

Chemotaxis transducers are specialized receptors that microorganisms use in order to sense the environment in directing their motility to favorable niches. The Escherichia coli transducers are models for studying the sensory and signaling events at the molecular level. Extensive studies in other organisms and the arrival of genomics has resulted in the accumulation of sequences of many transducer genes, but they are not fully understood. In silico analysis provides some assistance in classification of various transducers from different species and in predicting their function. All transducers contain two structural modules: a conserved C-terminal multidomain module, which is a signature element of the transducer superfamily, and a variable N-terminal module, which is responsible for the diversity within the superfamily. These structural modules have two distinct functions: the conserved C-terminal module is involved in signaling and adaptation, and the N-terminal module is involved in sensing various stimuli. Both C-terminal and N-terminal modules appear to be mobile genetic elements and subjects of duplication and lateral transfer. Although chemotaxis transducers are found exclusively in prokaryotic organisms that have some type of motility (flagellar, gliding or pili-based), several types of domains that are found in their N-terminal modules are also present in signal transduction proteins from eukaryotes, including humans. This indicates that basic principles of sensory transduction are conserved throughout the phylogenetic tree and that the chemotaxis transducer superfamily is a valuable source of novel sensory elements yet to be discovered.

Amino Acid Sequence↗
PubMed1999

In vivo methotrexate selection of murine hemopoietic cells transduced with a retroviral vector for Gaucher disease.

The studies described were performed to investigate whether in vivo selection of retrovirus-transduced hemopoietic cells is feasible starting from a low percentage of transduced hemopoietic stem cells (PHSCs). The vector used is an amphotropic bicistronic retroviral vector carrying a cDNA for human lysosomal glucocerebrosidase (hGC) for treatment of Gaucher disease and a methotrexate (MTX) resistant mutant cDNA encoding human dihydrofolate reductase (DHFR). We tested the effect of MTX selection in mice that were either myeloablated or not before infusion of transduced cells. In addition, we determined whether repeated administration of transduced bone marrow cells has an additional effect on the percentage of hGC expressing cells. The results obtained have shown that, in myeloablated mice transplanted once with transduced bone marrow and treated twice weekly with 10 mg/kg of MTX for a total of 6 months, a two- to three-fold increased numbers of hGC expressing cells could be detected in both peripheral blood and bone marrow as compared with non-MTX treated mice. In mice transplanted with transduced bone marrow once every 2 weeks for a total of four times, percentages of hGC expressing cells were not significantly increased as compared with mice transplanted once. In non-ablated mice neither MTX selection nor multiple infusions of transduced bone marrow resulted in detection of hGC expressing cells 6 months after transplantation, indicating that the success of in vivo selection using MTX is highly dependent on the ratio of transduced hemopoietic stem cells transplanted versus residing and untransduced stem cells.

Animals↗
PubMed1996

The primary structures of the Archaeon Halobacterium salinarium blue light receptor sensory rhodopsin II and its transducer, a methyl-accepting protein.

Recently, a large family of transducer proteins in the Archaeon Halobacterium salinarium was identified. On the basis of the comparison of the predicted structural domains of these transducers, three distinct subfamilies of transducers were proposed. Here we report isolation, complete gene sequences, and analysis of the encoded primary structures of transducer gene htrII, a member of family B, and its blue light receptor gene (sopII) of sensory rhodopsin II (SRII). The start codon ATG of the 714-bp sopII gene is one nucleotide beyond the termination codon TGA of the 2298-bp htrII gene. The deduced protein sequence of HtrII predicts a eubacterial chemotaxis transducer type with two hydrophobic membrane-spanning segments connecting sizable domains in the periplasm and cytoplasm. HtrII has a common feature with HtrI, the sensory rhodopsin I transducer; like HtrI, HtrII possesses a hydrophilic loop structure just after the second transmembrane segment. The C-terminal 299 residues (765 amino acid residues total) of HtrII show strong homology to the signaling and methylation domain of eubacterial transducer Tsr. The hydropathy plot of the primary structure of SRII indicates seven membrane-spanning alpha-helical segments, a characteristic feature of retinylidene proteins ("rhodopsins") from a widespread family of photoactive pigments. SRII shows high identity with SRI (42%), bacteriorhodopsin (BR) (32%), and halorhodopsin (24%). The crucial positions for retinal binding sites in these proteins are nearly identical, with the exception of Met-118 (numbering according to the mature BR sequence), which is replaced by Val in SRII. In BR, residues Asp-85 and Asp-96 are crucial in proton pumping. In SRII, the position corresponding to Asp-85 in BR is conserved, but the corresponding position of Asp-96 is replaced by an aromatic Tyr. Coexpression of the htrII and sopII genes restores SRII phototaxis to a mutant (Pho81) that contains a deletion in the htrI/sopI and insertion in htrII/sopII regions. This paper describes the first example that both HtrI and HtrII exist in the same halobacterial cell, confirming that different sensory rhodopsins SRI and SRII in the same organism have their own distinct transducers.

Amino Acid Sequence↗
PubMed1999

The specificity of interaction of archaeal transducers with their cognate sensory rhodopsins is determined by their transmembrane helices.

Chimeras of the Halobacterium salinarum transducers HtrI and HtrII were constructed to study the structural determinants for their specific interaction with the phototaxis receptors sensory rhodopsins I and II (SRI and SRII), respectively. Interaction of receptors and transducers was assessed by two criteria: phototaxis responses by the cells and transducer-modulation of receptor photochemical reaction kinetics in membranes. Coexpression of HtrI with SRII or HtrII with SRI did not result in interaction by either criterion. Each receptor was coexpressed with chimeric transducers in which various domains of the two transducers were interchanged. The results show that the presence of the two transmembrane helices of HtrI in a chimera is necessary and sufficient for functional transducer complexation with SRI, i.e., for wild-type SRI photoreactions and attractant and 2-photon repellent phototaxis responses. Additionally, a previously demonstrated chaperone-like facilitation of SRI folding or stability by HtrI was shown to depend only on the two transmembrane helices of HtrI in chimeric transducers. Similarly, the two transmembrane helices of HtrII specify interaction with the repellent receptor SRII according to motility analysis and laser-flash spectroscopy. The results support a model in which the membrane domains of the receptor/transducer complexes, consisting of the seven helices of the receptor interacting with the four-helix bundle of the transducer dimer, produce SRI- and SRII-specific signals to the flagellar motor by means of interchangeable cytoplasmic domains.

Archaeal Proteins↗
PubMed1980

Studies on the mechanism of transduction by bacteriophage phi gamma. II. Formation of transducing elements.

The formation of the transducing elements (TE) of bacteriophage phi gamma, analyzed in lysogens of the thermo-inducible derivative phi gamma hyI, has been found to parallel the formation of plaque-forming particles with a frequency of 2 X 10(-2) TE/PFU, but is mor sensitive to temperature and ti UV. Deletion of one of the prophage termini (attR) prevents normal excision and formation of plaque-forming particles, but does not affect the formation of transducing elements, which arise at a rate of nearly 10(-1) TE per induced bacterium. Transducing elements, would be formed by in situ encapsulation of a hybrid segment from a specidic point in the induced prophage, possibly the presumed packaging initiation site of the normal phage genome, before excision of the latter has occurred. Analysis of the mechanism of transduction to partly heterologous lysogens has revealed the participation of a co-infecting genome arranged in a linear fashion and has given evidence for a permutation in the sequence of transducing and nontransducing genomes. The data re consistent with a mechanism of encapsidation distinct from the Ter system even for hybrids inheriting part of the phi 80 genome, but endowed with the property to form transducing elements like those of phi gamma. Upon infection, transducing elements are formed after one cycle of lytic development with the same characteristics as those resulting from induction, but with a frequency 50 to 100 times lower. This process is dependent on the efficiency of Int promoted recombination. Superinfection experiments performed under conditions preventing Int promoted recombination reveal that any superinfecting phi gamma can promote the formation of transducing particles, depending on the presence within the host prophage of a site from which transducing genome packaging initiates.

Chromosome Mapping↗