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At least 523 records · Page 29Linked to original sources
PubMed2001

Melon bacterial artificial chromosome (BAC) library construction using improved methods and identification of clones linked to the locus conferring resistance to melon Fusarium wilt (Fom-2).

Utilizing improved methods, two bacterial artificial chromosome (BAC) libraries were constructed for the multidisease-resistant line of melon MR-1. The HindIII library consists of 177 microtiter plates in a 384-well format, while the EcoRI library consists of 222 microtiter plates. Approximately 95.6% of the HindIII library clones contain nuclear DNA inserts with an average size of 118 kb, providing a coverage of 15.4 genome equivalents. Similarly, 96% of the EcoRI library clones contain nuclear DNA inserts with an average size of 114 kb, providing a coverage of 18.7 genome equivalents. Both libraries were evaluated for contamination with high-copy vector, empty pIndigoBac536 vector, and organellar DNA sequences. High-density filters were screened with two genetic markers FM and AM that co-segregate with Fom-2, a gene conferring resistance to races 0 and 1 of Fusarium wilt. Fourteen and 18 candidate BAC clones were identified for the FM and AM probes, respectively, from the HindIII library, while 34 were identified for the AM probe from filters A, B, and C of the EcoRI library.

Chromosomes, Artificial, Bacterial↗
PubMed2002

[Expression and identification of phage display library for Fab fragments of colorectal cancer-related antibodies].

OBJECTIVE: To express the original human Fab antibody phage display library with positive recombined bacterium XL1-blue-Pcomb3 and identify its specific binding activity with colorectal cancer cells in vitro after screening with human colorectal cancer-related antigens. METHOD: The recombination rate of Fd fragment of the heavy chain and insertion of kappa chain of the antibodies was determined with PCR, and the original Fab library was expressed. The antigens were extracted from 3 sensitized colorectal cancer tissues previously used for construction of the original Fab library and from 13 non-sensitized colorectal cancer tissues, along with the antigens from LoVo, HT-29 and LS-174T cells cultured in vitro. The original Fab antibody library was screened with the 3 groups of mixed antigens derived in preceding procedure and 3 tertiary Fab antibody libraries were obtained, which were then mixed in equal volume for subsequent tests of binding activity with human colorectal cancer tissues and cells in vitro using enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining. Specimens of gastric and esophageal carcinomas and normal intestinal mucosa, together with liver cancer cells and gastric cancer cells were utilized as control. RESULT: The recombination rate of Fd and kappa chain were 40 % and 70 % respectively, and the rate of their simultaneous insertion into Pcomb3 vector was 28%. The capacity of library for Fab fragment genes was 2.1x10(6), and the original antibody libraries screened with the 3 groups of mixed antigens were enriched to varied degrees, which all displayed relatively specific binding activity with human colorectal cancer tissue and cells in vitro. CONCLUSION: Colorectal cancer-related antibody Fab fragments are obtained through screening phage display library, which show relatively specific binding activity with human colorectal cancer tissues and cells.

Antibodies, Neoplasm↗
PubMed2003

[Construction,identification and amplification of a yeast two-hybrid random cycle peptide library].

AIM: To construct a cycle peptide library composed of 16 random amino acids with yeast two-hybrid system. METHODS: Random oligonucleotides encoding 16-mer peptides were designed and synthesized artificially, and then were amplified by PCR. The amplified products were digested with BamH I and EcoR I and cloned into yeast expression plasmid pGADT(7) GH to construct the cycle library plasmids pGADT(7) GH-RP Then the number of different recombinants and the randomness of the library were tested, and the cycle peptide library plasmids were amplified, extracted and purified. RESULTS: A random cycle peptide library with 1.28 x10(7) different recombinant clones was obtained. No significant difference was found between amino acid distribution in the cycle peptide library and the expected frequency. CONCLUSION: The random cycle peptide library has been successfully constructed. And a lot of cycle peptide library plasmids with high purity were obtained.

Amino Acid Sequence↗
PubMed2005

Computational design of combinatorial peptide library for modulating protein-protein interactions.

Screening phage-displayed combinatorial peptide library is an effective approach for discovery of peptide modulators for protein-protein interactions. However, as peptide length increases, the chance of finding active peptides in a finite size library diminishes. To increase the likelihood of finding peptides that bind to a protein, we develop statistical potential for computational construction of biased combinatorial antibody-like peptide libraries. Based on the alpha shapes of antibody-antigen complexes, we developed an empirical pair potential for antigen-antibody interactions that depends on local packing. We validate this potential and show that it can successfully discriminate the native interface peptides from a simulated library of 10,000 random peptides for 34 antigen-antibody complexes. In addition, we show that it can successfully recognize the native binding surface patch among all possible surface patches taken from either the antibody or the antigen for seven antibody-antigen protein complexes contained in the CAPRI (Critical Assessment of Predicted Interactions) dataset. We then develop a Weighted Amino Acid Residue sequence Generator (WAARG) for design of biased peptide library. When compared with a random peptide library, WAARG libraries contain more native-like binding peptides at a significantly smaller size. Our method can be used to construct peptide library for screening of antibody variants with improved specificity and affinity to a target antigen. It can also be used for screening of antibody-like antagonist peptides modulating other protein-protein interactions.

Amino Acid Sequence↗
PubMed2005

[Construction of a cDNA library of Alternaria sp. strain JH505 using Gateway technology].

A cDNA library was constructed based on the Gateway system that is a method to construct a high-quality cDNA library without the use of restriction enzyme for cloning. This is the first report that the Gateway system is used to construct a cDNA library for Alternaria sp.. The entry cDNA library was constructed in this report has a high titer of 1 x 10(7) cfu/mL and contain a total clones of 9 x 10(7) cfu, with an average inserts size of about 1510bp. In order to screen for a gene encoding the plant activator protein from library using an antiserum, the entry cDNA library was transferred into Gateway destination vector to create an expression library. The expression library show a high titer of 1.58 x 10(6) cfu/mL and contains a total clones of 6.32 x 10(6) cfu, with an average inserts size of 1680bp.

Alternaria↗
PubMed2006

[Construction and identification of cDNA library from peripheral blood lymphocytes of Tibetans in Qinghai-Tibet plateau].

AIM: To construct the full length cDNA library of the peripheral blood lymphocytes in Tibetans and to identify the quality of the library. METHODS: The lymphocytes were separated from healthy Tibetan fresh blood, and total RNA was extracted. The full length cDNA library was constructed by SMART (switching mechanism at 5' end of RNA transcript) technique. The unamplified library was titered, and the percentage of recombinant clones were determined. The library was amplified, and the size of the inserts was identified by PCR. RESULTS: The titer of the constructed cDNA library was 1.8x10(9) pfu/L with the percentage of recombinant clones above 98%, and that of the amplified library was 8x10(12) pfu/L. The insert's size ranged from 0.75 to 2.0 kb with the average length of 1.35 kb. CONCLUSION: The constructed cDNA library of peripheral blood lymphocytes of Tibetans has excellent quality, and it lays a solid founclation for further screening and cloning hypoxia-related genes of Tibetans living at high altitudes.

Asian People↗
PubMed1991

Hybridization selections, using immobilized driver DNA, to enrich for clones unique to a library.

We have developed a technique which allowed us to isolate sex-specific repeats of chickens (Gallus g. domesticus) from a genomic library originally containing one sex-specific repeat clone per 300 clones. Using this plus/minus selection method, we were able to enrich a sub-library in which the sex-specific repetitive clones made up over half of the population (170-fold increase in representation). This enrichment technique used hybridization kinetics to collect clones which are bound (plus selection) or not bound (minus selection) to their respective driver DNAs immobilized on nitrocellulose paper. Plus selections enriched for repetitive sequences and removed from the library most unique sequences as well as vectors containing no inserted sequences. These plus-selected sub-libraries were enriched for repetitive clones, yet still contained sequences representing as little as 10(-4) of the genome. Minus selection removed repetitive sequences shared between the driver and the library. When a plus-selected sub-library was minus selected, the doubly selected sub-library was enriched in repetitive sequences present in the library but not the minus driver.

Animals↗
PubMed1970

Kansas Regional Medical Program Library Services.

The Kansas Regional Medical Program Office for Library Services was developed to link the medical library resources and to make them available to health-related personnel throughout Kansas. Library offices have been established at the Central Kansas Medical Center, Great Bend, Stormont Medical Library, Topeka, and at the Wichita State University, Wichita. The main office, located at the Clendening Medical Library, University of Kansas Medical Center, Kansas City, provides medical library services to those three offices, the other medical libraries in Kansas, and to the health professionals in the state who do not have medical library resources available. Reference requests are accepted via a twenty-four-hour telephone service from medical librarians and from individual health professionals.

Communication↗
PubMed1969

Planning for a library system: Connecticut Regional Medical Program.

A formal medical library system is developing nationally to improve library service, but not all users or even all librarians are alert to the need. The main stimulus seems to come from federal money and from leaders at the top, rather than from the small local library and its user, yet progress depends on participation at all levels. Planning for a state-wide medical library system as part of the Connecticut Regional Medical Program began with a survey of the state's medical library resources, which led to a grant request for operating funds to strengthen reference and inter-library loan service in Connecticut and to begin a training and consultation program for medical librarians in the state. These activities are intended to expand and intensify in Connecticut those back-up services provided for all of New England by the New England regional medical library service at the Countway Library in Boston and also are related to the other Regional Medical Program activities planned for Connecticut.

Connecticut↗
PubMed1978

Departmental libraries: curse or blessing?

Communication patterns and working relationships between large health sciences libraries and departmental libraries are examined and discussed. The results of a telephone survey to obtain information on the experiences of twenty-one large New York metropolitan area health sciences libraries with their departmental libraries are reported. Discussion includes the experiences of Columbia University's Augustus Long Health Sciences Library with departmental libraries within the Columbia/Presbyterian Medical Center (C/PMC). A mediated survey of C/PMC departmental libraries and over thirty years of cooperation with some of these departments are reported. The potential detrimental effects of departmental libraries are acknowledged, but the substantial positive benefits of cooperation and ways to achieve this cooperation are emphasized.

Attitude↗
PubMed1995

A descriptive study of ninety-two hospital libraries in Mexico.

This work reports on the existing situation of ninety-two hospital libraries, located at Mexico's Social Security Institute (IMSS). A descriptive, systems approach was used to explore the physical structure and space of the libraries, staff, furniture, equipment, collections, services, organization and management, and users. Structured interviews were applied to each hospital library, and a questionnaire was used as a tool to collect data. A "control" system, designed to measure the status of each library, was applied through the assignment of values to several indicators, derived from IMSS policy manuals. This procedure helped to identify which library was "more" or "less" adequate to IMSS standards. Within the rank of 10 to 1 (10 = optimum; 1 = minimum), the mean rank of IMSS hospital libraries was 6. The major deficiencies found were those related to furniture (ranked 4) and services (ranked 5) and the lack of library professionals found in 92% of the libraries.

Academies and Institutes↗
PubMed2003

Current issues in the design of academic health sciences libraries: findings from three recent facility projects.

Planning a new health sciences library at the beginning of the twenty-first century is a tremendous challenge. Technology has radically changed the way libraries function in an academic environment and the services they provide. Some individuals question whether the library as place will continue to exist as information becomes increasingly available electronically. To understand how libraries resolve programming and building design issues, visits were made to three academic health sciences libraries that have had significant renovation or completed new construction. The information gathered will be valuable for planning a new library for the University of Colorado Health Sciences Center and may assist other health sciences librarians as they plan future library buildings.

Colorado↗
PubMed1977

Improving library effectiveness through a sociophysical analysis.

A library's environment as an inviting place to study, read, and relax is an important function often overlooked or avoided. A nursing school library was underutilized and had an unacceptable of lost materials. A study undertaken to determine the setting's impact on all the library's intended functions showed that the physical environment seriously hampered many activities. A change in the library's arrangement offered users a more functional area for all types of library activities. Statistics show that the library is now more fully utilized, with an impressive decline in losses. The majority of students and faculty report more positive feelings toward the library and the librarian.

Environment↗
PubMed1964

THE DEVELOPMENT OF INTERLIBRARY LOAN SERVICE IN JAPANESE MEDICAL LIBRARIES.

An interlibrary loan system in Japan was established by the Japanese Medical Library Association in 1927. Since then the members of the Association have increased from five to forty-eight-all forty-six medical school libraries and two dental school libraries. The Association's service has been enlarged, particularly since World War II. The number of interlibrary loans among member libraries has increased greatly, especially since 1954, thanks to the development of union lists and photoduplication service. Today, more than 80 percent of the requests are filled with photocopied materials. On the other hand, the growth of medical literature has made interlibrary cooperation very necessary, especially internationally. An agreement was made in 1948 concerning photoduplication service between the Japanese Medical Library Association and the National Library of Medicine.

Computer Systems↗
PubMed2005

Bacterial community composition in different sediments from the Eastern Mediterranean Sea: a comparison of four 16S ribosomal DNA clone libraries.

The regional variability of sediment bacterial community composition and diversity was studied by comparative analysis of four large 16S ribosomal DNA (rDNA) clone libraries from sediments in different regions of the Eastern Mediterranean Sea (Thermaikos Gulf, Cretan Sea, and South lonian Sea). Amplified rDNA restriction analysis of 664 clones from the libraries indicate that the rDNA richness and evenness was high: for example, a near-1:1 relationship among screened clones and number of unique restriction patterns when up to 190 clones were screened for each library. Phylogenetic analysis of 207 bacterial 16S rDNA sequences from the sediment libraries demonstrated that Gamma-, Delta-, and Alphaproteobacteria, Holophaga/Acidobacteria, Planctomycetales, Actinobacteria, Bacteroidetes, and Verrucomicrobia were represented in all four libraries. A few clones also grouped with the Betaproteobacteria, Nitrospirae, Spirochaetales, Chlamydiae, Firmicutes, and candidate division OPl 1. The abundance of sequences affiliated with Gammaproteobacteria was higher in libraries from shallow sediments in the Thermaikos Gulf (30 m) and the Cretan Sea (100 m) compared to the deeper South Ionian station (2790 m). Most sequences in the four sediment libraries clustered with uncultured 16S rDNA phylotypes from marine habitats, and many of the closest matches were clones from hydrocarbon seeps, benzene-mineralizing consortia, sulfate reducers, sulk oxidizers, and ammonia oxidizers. LIBSHUFF statistics of 16S rDNA gene sequences from the four libraries revealed major differences, indicating either a very high richness in the sediment bacterial communities or considerable variability in bacterial community composition among regions, or both.

Bacteria↗
PubMed2002

Evaluation of lipophilicity and antitumour activity of parallel carboxamide libraries.

Searching for molecules possessing antitumour activity, a parallel molecule library of aromatic carboxamides has been designed and synthesised. This work resulted in a "thiophene" sub-library containing a thiophene core and of a "furoyl" sub-library with a furoyl core, respectively. In both sub-libraries substitutions were carried out with six different groups resulting in six pairs of compounds differing in only the heteroatom of aromatic ring of the cores. To study the importance of the type of cores and the specific substitutions in relation to their lipophilicity and antitumour activity, lipophilicity of carboxamides was determined by chromatographical data (log k') and by software calculated parameters (CLOGP). Pairs of compounds were tested for their ability to inhibit the proliferation of the A431 cells by MTT assay. The isosteric molecule pairs were successfully separated. Our results showed that the experimentally determined (log k') and the calculated (CLOGP) lipophilicity parameters correlated well with each other. Furthermore, lipophilicity values of the thiophene sub-library were always higher than those in the furoyl sub-library. Moreover, compounds of the thiophene sub-library were more active than their respective furoyl pairs in our MTT antiproliferative assay. From these observations we can conclude that the higher the lipophilicity values the higher the antitumour activity of the carboxamides synthesised. Therefore, determination of lipophilicity by measuring the log k' or by calculating the CLOGP values of the carboxamide sub-libraries may help to predict their biological activities.

Amides↗
PubMed2005

High-throughput solution-based medicinal library screening against human serum albumin.

High-throughput screening of combinatorial libraries has evolved from studying large diverse libraries to analyzing small, structurally similar, focused libraries. This paradigm shift has generated a need for rapid screening technologies to screen both diverse and focused libraries in a simple, efficient, and inexpensive manner. We have proactively addressed these needs by developing a high-throughput, solution-based method combining size exclusion (SEC), two-dimensional liquid chromatography (2-D LC), and mass spectrometry (MS) for determining the relative binding of drug candidates in small, focused medicinal libraries against human serum albumin (HSA). Two types of libraries were used to evaluate the performance of the system. The first consisted of five diverse ligands with a wide range of hydrophobicities and whose association constants to HSA cover 3 orders of magnitude. A beta-lactam library composed of structurally similar compounds was used to further confirm the validity of the methodology. The ability to distinguish site-specific interactions of drugs competing for individual domains of the HSA receptor is also demonstrated. Comparison of chromatographic profiles of the library components before and after incubation with the receptor using multiple reaction monitoring allowed a ranking of the ligands according to their relative binding affinities. The observed rankings correlate closely with literature values of the association constants between the respective ligands and HSA. This simple, rugged methodology can screen a wide spectrum of chemical entities from combinatorial mixtures in less than 6 min.

Acecainide↗
PubMedDate not supplied

Parallel microwave-assisted library of imidazothiazol-3-ones and imidazothiazin-4-ones.

A methodology for the generation of a microwave-assisted parallel library and its conversion into a second library is described. A 24-membered library of substituted 4(5)-sulfanyl-1H-imidazoles was generated and subsequently converted into a second library of bicyclic imidazo[5,1-b]thiazol-3-ones and imidazo[5,1-b]thiazin-4-ones. The first library was generated using a three-component reaction and transformed into a daughter library with a polymer-supported coupling agent. The procedure involved the use of an array of expandable reaction vessels, which can accommodate pressure buildup due to microwave heating without loss of volatile solvents or reagents. Library generation time for each library was 16 min.

Journal Article↗