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PubMed1999

Health sciences library building projects, 1998 survey.

Twenty-eight health sciences library building projects are briefly described, including twelve new buildings and sixteen additions, remodelings, and renovations. The libraries range in size from 2,144 square feet to 190,000 gross square feet. Twelve libraries are described in detail. These include three hospital libraries, one information center sponsored by ten institutions, and eight academic health sciences libraries.

Communications Media↗
PubMed2001

[The construction of rapid amplification of cDNA ends cDNA libraries from human fetal bone and joint].

OBJECTIVE: To construct rapid amplification cDNA ends(RACE) cDNA libraries from human fetal bone and joint and provide resources for isolation of bone- and joint-specific development-related genes. METHODS: Total RNA of bone and joint were extracted with the modified single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. The double-stranded end-blunted cDNA were synthesized using TaKaRa's cDNA synthesis kit and ligated to cassette adaptors. All of the cDNA molecules were amplified by a pair of common primers. RESULTS: A protocol for RACE cDNA library construction from bone and joint was established and two RACE cDNA libraries from human fetal bone and joint were successfully constructed. CONCLUSION: The protocol of RACE cDNA library construction from limited materials proved to be simple and efficient and the library was suitable for RACE to isolate tissue-specific genes.

Bone and Bones↗
PubMed1975

Mobilization of duplicates in a Regional Medical Library Program.

An overabundance of duplicate journals without an efficient and economical method of distribution caused one library's staff to reassess traditional methods of dispersal. A simplified form for listing duplicates was devised. In conjunction with the Regional Medical Library Program and the extension program, lists of duplicates were distributed to hospital and clinical libraries. These libraries selected materials to strengthen their ability to fill information needs at the local level and to conserve RMLP support for esoteric and expensive materials. In a two-year period, 86,000 individual pieces were distributed. Some lessening of interlibrary loan requests from heavy users was documented. In an evaluation survey users expressed satisfaction with the program. The successful use of the duplicate program will lead to a further experiment--the library will attempt to fill interlibrary loan requests for common journals with hard copy rather than photocopy in a cost and time reduction effort.

Book Collecting↗
PubMed2001

A bacterial artificial chromosome (BAC) library of Malus floribunda 821 and contig construction for positional cloning of the apple scab resistance gene Vf.

The apple scab resistance gene Vf, originating from the wild species Malus floribunda 821, has been incorporated into a wide variety of apple cultivars through a classical breeding program. With the aim of isolating the Vf gene, a bacterial artificial chromosome (BAC) library consisting of 31 584 clones has been constructed from M. floribunda 821. From the analysis of 88 randomly selected BAC clones, the average insert size is estimated at 125 kb. If it is assumed that the genome size of M. floribunda 821 is 769 Mb/haploid, the library represents about 5x haploid genome equivalents. This provides a 99% probability of finding any specific sequence from this library. PCR-based screening of the library has been carried out using eight random genomic sequence-characterized amplified regions (SCARs), chloroplast- and mitochondria-specific SCARs, and 13 high-density Vf-linked SCAR markers. An average of five positive BAC clones per random SCAR has been obtained, whereas less than 1% of BAC clones are derived from the chloroplast or mitochondrial genomes. Most BAC clones identified with Vf-linked SCAR markers are physically linked. Three BAC contigs along the Vf region have been obtained by assembling physically linked BAC clones based on their fingerprints. The overlapping relatedness of BAC clones has been further confirmed by cytogenetic mapping using fiber fluorescence in situ hybridization (fiber-FISH). The M. floribunda 821 BAC library provides a valuable genetic resource not only for map-based cloning of the Vf gene, but also for finding many other important genes for improving the cultivated apple.

Chromosomes, Artificial, Bacterial↗
PubMed2000

[Selection and characterization of human antibodies against vascular endothelial growth factor from human phage display antibody library].

OBJECTIVE: To select anti-VEGF (vascular endothelial growth factor) antibodies (Fab fragments) from human phage display antibody library and to identify their specification and activities. METHODS: Human immunoglobulin heavy chain and light chain genes were separately amplified by RT-PCR from human peripheral lymphocytes using family specific primers and signal sequences of immunoglobulin. Human antibody library was constructed by phage display technology, and phage Fab antibodies to VEGF were screened from this library. ELISA, Western blot and (3)H-thymidine incorporation assays were used for the specification and neutralization activities of these Fab antibodies. Sequencing analysis was carried out for further identification of the antibodies. RESULTS: The repertoire of human phage display Fab library was 1.5 x 10(8). After 4 round panning with VEGF(121), 280 clones were checked for their binding activities with ELISA and 12 clones could bind to VEGF(121) specifically. Western blot demonstrated that bacterially expressed soluble Fab could specifically recognize VEGF. Results of (3)H- thymidine incorporation showed that one clone of soluble Fab could neutralize the mitogenic activity of VEGF(165) on HUVEC. Sequencing analysis showed that the obtained V(H) gene belonged to human VH6 subgroup and the light chain was VJC rearranged human VL4 gene. CONCLUSIONS: Human anti-VEGF antibodies can be obtained from human phage display library, which provides a basis for preparation of high affinity human anti-VEGF monoclonal antibodies through antibody engineer technique.

Antibodies↗
PubMed2001

[Construction of cDNA library of Eimeria tenella sporulated oocysts].

A lambda ZAP express cDNA library was constructed using mRNA from Eimeria tenella sporulated oocysts. Total RNA was isolated by the TRIzol from Eimeria tenella sporulated oocysts, mRNA was further purified through oligo(dT)-cellulose columns. The first-strand cDNA was synthesized by using MMLV reverse transcriptase with oligo(dT)18 primers containing Xho I restriction site. After the second strand cDNA replacement synthesized, the uneven termini of the double-stranded cDNA were filled in with cloned Pfu DNA polymerase and EcoR I adapters were ligated to the blunt ends. Then the double-strand cDNA was digested with Xho I restriction enzyme. The fragments of 0.5 kb-4 kb were collected by agarose gel fraction method. After ligation of the cDNA with the lambda ZAP Express vector, the cDNA library was packaged using Gigapack III Gold Packaging extract. According to the phage plaques bright selection, the cDNA library contained 6 x 10(6) clones and the titer of the amplified library was 1 x 10(11) pfu/mL. By using PCR identification, the cDNA library contained approximately 96% recombinant phages.

Animals↗
PubMed1998

[Construction and screening of type 1 human immunodeficiency virus specific phage antibodies combinatorial library].

UNLABELLED: Human monoclonal antibodies to type 1 immunodeficiency virus (HIV-1) gp120 were generated from phage antibody combinatorial library. METHODS: The human immunoglobulin heavy chain Fd and light chain k genes were amplified by half-nested PCR from PBMC of patient infected with HIV. Phage antibody combinatorial library was constructed with the Fd and k chain genes using Pcomb3 as vector. The affinity selection and ELISA were adopted for generating specific phage antibodies. Partial DNA of a positive clone was sequenced and its soluble Fab was expressed in E coli. HIV-1 specific phage antibodies combinatorial library were constructed using the Fd and k genes and Pcomb3 vector. The library capacity was about 1.95 x 10(7). The specific phage antibodies were highly enriched after three rounds of biopanning selection against HIV-1 gp120 and 32% positive clones were detected by ELISA screening. DNA fragment coding for CH1 and CL derived from a positive clone was sequenced and its product was successfully expressed as soluble Fab which was specific for HIV-1 gp120. The HIV-1 specific phage antibody combinatorial library, and human monoclonal antibodies to HIV-1 gp120 have been used as tools for screening of neutralizing antibody to HIV-1, and the methods seem to be very crucial and applicable.

Amino Acid Sequence↗
PubMed2002

[Construction and characterization of the cDNA library from Schistosoma japonicum cercariae].

OBJECTIVE: To construct a cDNA library from Schistosoma japonicum cercariae. METHODS: Total RNA was extracted from 500,000 S. japonicum cercariae and used to synthesize the double-stranded cDNA and cloned into the TriplEx2 vector using "SMART cDNA Construction Kit". RESULTS: The primary titer of the constructed cDNA library is 1.8 x 10(7) pfu/ml and the titer of amplified library is 2.5 x 10(10) pfu/ml. The average size of inserts is 1.075 kb. The recombinant efficiency is 94.4%. The full length cDNA of S. japonicum TPI and JF-2 genes was successfully amplified from the library. CONCLUSION: A cDNA library of S. japonicum cercariae has been constructed successfully.

Animals↗
PubMed2003

[Rapid construction and identification of full-length cDNA library of human glioma tissues].

OBJECTIVE: To explore a method for rapid construction of a full-length cDNA library of human glioma tissues using switching mechanism at 5' end of RNA transcript (SMART). METHODS: The total RNA was extracted from several samples of human glioma tissues and the mRNA was subsequently separated. Multiple mRNA samples were mixed to be used as the template for the first-strand cDNA synthesis. The CDS /3' PCR primer (containing Sfi IB site) was used in the first-strand reaction, and the SMART IV Oligo(dT) (containing Sfi A site) served as the short, extended template at the 5' end of the mRNA. With the above two primers, the primer-extension step generated full-length double-strand cDNA, which was digested by Sfi I restriction enzyme and ligated to the Sfi I A & B -digested lambdaTriplEx2 vector. The ligated vector was then packaged by lambda packaging extract for the final construction of the cDNA library. RESULTS: The unamplified human glioma cDNA library consisted of 2.4x10(6) independent clones with a recombination rate of 100%. The titer of the amplified cDNA library was 4.5x10(9) pfu/ml, and the average exogenous inserts of the recombinants was 1.2 kb in length. CONCLUSION: A high-quality full-length cDNA library of human gliomas was constructed successfully, which may facilitate further study of the screening and cloning of new tumor suppressor genes and tissue-specific genes of human glioma.

DNA, Complementary↗
PubMed2003

A large-insert porcine library with sevenfold genome coverage: a tool for positional cloning of candidate genes for major quantitative traits.

A porcine genomic bacterial artificial chromosome (BAC) library was constructed by cloning partial EcoRI-digested high-molecular-weight DNA from a Korean native boar into the EcoRI site of the pBACe3.6 vector. The library consists of about 165,000 clones with an average insert size of 125 kb, representing about seven genome equivalents of coverage. About 130,000 clones (corresponding to fivefold genome coverage) were arrayed in 14 superpools which were organized as four dimensional pools. The library was further characterized by PCR screening of 38 microsatellite probes. An average of 4.84 positive clones were selected per marker. This indicates that the library is unbiased and will be useful for initiating fine scale physical mapping of major QTL in pigs. The library is being used to isolate specific clones by screening with type I and type II marker clones located in the QTL region affecting intramuscular fat content on SSC6.

Animals↗
PubMed2003

[Construction of forward and reverse subtracted cDNA libraries between muscle tissue of Meishan and Landrace pigs].

Using suppression subtractive hybridization (SSH) technique, forward and reverse subtracted cDNA libraries were constructed between Longissimus muscles from Meishan and Landrace pigs. A housekeeping gene, G3PDH, was used to estimate the efficiency of subtractive cDNA. In two cDNA libraries, G3PDH was subtracted very efficiently at appropriate 2(10) and 2(5) folds, respectively, indicating that some differentially expressed genes were also enriched at the same folds and the two subtractive cDNA libraries were very successful. A total of 709 and 673 positive clones were isolated from forward and reverse subtracted cDNA libraries, respectively. Analysis of PCR showed that most of all plasmids in the clones contained 150-750 bp inserts. The construction of subtractive cDNA libraries between muscle tissue from different pig breeds laid solid foundations for isolating and identifying the genes determining muscle growth and meat quality, which will be important to understand the mechanism of muscle growth, determination of meat quality and practice of molecular breeding.

Animals↗
PubMed2003

[Construction of subtracted cDNA library in human Jurkat T cell line induced by arsenic trioxide in vitro].

OBJECTIVE: To understand the differentially expressed genes in human T lymphocytes induced by arsenic trioxide (As(2)O(3)) and to explore mechanism of its immunotoxicity and immune suppression. METHODS: Human Jurkat T cell line was treated by arsenic trioxide (5 micromol/L, 24 h) in vitro, as a sample model. Then, the differentially expressed genes were cloned and the subtractive cDNA library from Jurkat T cell line was constructed by suppression subtractive hybridization (SSH). Polymerase chain reaction (PCR) and sequencing techniques were applied to identify positive clones. RESULTS: The forward subtracted cDNA library contained differentially expressed genes from Jurkat T cell line induced by arsenic trioxide was constructed, including 29 different gene fragments and only replicated one in the subtracted cDNA library identified by PCR and sequencing analysis. These gene sequences were 95%-100% analogous to the genes in public database (GenBank/EMBL). The cDNA library contained oxidative metabolic genes in mitochondria (triose phosphate dehydrogenase, NADH4, pyrophosphate synthase, 16S rRNA ribosome, succinate-CoA ligase and ATP synthase 6); transcriptional and translation genes poly (A) binding protein, t-RNA-guanine transglycoslase, ribosomal protein L23, ribosomal protein S15A, eukaryotic translation initiation factor 3, Rab interaction protein 5, splicing factor-arginine serine rich 5, and ADP-ribosylation factor-like 6 interacting protein), oxide stress related genes (ferritin high chain and high-mobility group protein 2); protein activating and signaling pathway related genes (casein kinase, serine kinase 2 and phosphatidylinositol-four-phosphate adaptor protein-1-associated protein); cell differentiation and apoptosis associated genes (NB4 cell apoptosis related protein and myeloid differentiation primary response protein) and five genes with unknown function (KIAA0092, CGI-147protein, GCI-35, nucleolar phosphoprotein Nopp34 and Mus muscular partial mRNA for hypothetical protein), as well as a novel gene unmatched to the sequence in GenBank. CONCLUSIONS: The forward subtracted cDNA library contained differentially expressed genes from Jurkat T cell line induced by arsenic trioxide was successfully constructed. And, genes not involved in previous research on arsenic were found. Results of analysis for these genetic function suggested that there should be many genes involved in process of T lymphocytes apoptosis or injury induced by arsenic trioxide and that there should still be many genes associated with arsenic that were not reported in the past.

Antineoplastic Agents↗
PubMed2002

New genomic resources for the honey bee(Apis mellifera L.): development of a deep-coverage BAC library and a preliminary STC database.

We have constructed a bacterial artificial chromosome (BAC) library for a European honey bee strain using the cloning enzyme HindIII in order to develop resources for structural genomics research. The library contains 36,864 clones (ninety-six 384-well plates). A random sampling of 247 clones indicated an average insert size of 113 kb (range = 27 to 213 kb) and 2% empty vectors. Based on an estimated genome size of 270 Mb, this library provides approximately 15 haploid genome equivalents, allowing >99% probability of recovering any specific sequence of interest. High-density colony filters were gridded robotically using a Genetix Q-BOT in a 4 x 4 double-spotted array on 22.5-cm2 filters. Screening of the library with four mapped honey bee genomic clones and two bee cDNA probes identified an average of 21 positive signals per probe, with a range of 7-38 positive signals per probe. An additional screening was performed with nine aphid gene fragments and one Drosophila gene fragment resulting in seven of the nine aphid probes and the Drosophila probe producing positive signals with a range of 1 to 122 positive signals per probe (average of 45). To evaluate the utility of the library for sequence tagged connector analysis, 1152 BAC clones were end sequenced in both forward and reverse directions, giving a total of 2061 successful reads of high quality. End sequences were queried against SWISS-PROT, insect genomic sequence GSS, insect EST, and insect transposable element databases. Results in spreadsheet format from these searches are publicly available at the Clemson University Genomics Institute (CUGI) website in a searchable format (http://www.genome.clemson.edu/projects/stc/bee/AM__Ba/).

Animals↗
PubMed2003

[Construction and characterization of anti-DOTA-Y phage antibody library].

AIM: To construct a anti-dodecane-tertraacetic acid-yttrium(DOTA-Y) immune Fab phage antibody library. METHODS: BALB/c were immunized with BSA-Y-DOTA which was prepared by DOTA-conjugated BSA and chelated with Y. After determination of anti-serum, total RNA was extracted from splenic lymphocytes of immuned mice. The heavy chain Fd and light chain Kappa genes repertoires of immunoglobulin were amplified respectively by RT-PCR, and then the amplified products were cloned into the reconstructive phage vector pComb3M to construct anti-DOTA-Y Fab antibody. And then, the recombination rate, diversity and display of Fab antibody library were identified by restriction endonuclease digestion, DNA sequencing and ELISA. RESULTS: BSA-Y-DOTA was prepared successfully, and a higher titer of immune sera was achieved. The amplified gene fragments of Fd and Kappa chain by RT-PCR were correct and the length was with about 650 bp, and were inserted exactly. The sink size of Fab phage display library reached 8 x 10(7), the re-combination rate was about 90%, and it possesed great diversity. In addition, ELISA detection showed that there was Fab expression on the phage library. CONCLUSION: An immune Fab phage antibody library of DOTA-Y has been constructed successfully, which lays a solid foundation for screening specific anti-DOTA-Y antibody.

Animals↗
PubMed1992

The medical libraries of Vietnam--a service in transition.

The medical libraries of Vietnam maintain high profiles within their institutions and are recognized by health care professionals and administrators as an important part of the health care system. Despite the multitude of problems in providing even a minimal level of medical library services, librarians, clinicians, and researchers nevertheless are determined that enhanced services be made available. Currently, services can be described as basic and unsophisticated, yet viable and surprisingly well organized. The lack of hard western currency required to buy materials and the lack of library technology will be major obstacles to improving information services. Vietnam, like many developing nations, is about to enter a period of technological upheaval, which ultimately will result in a transition from the traditional library limited by walls to a national resource that will rely increasingly on electronic access to international knowledge networks. Technology such as CD-ROM, Integrated Services Digital Network (ISDN), and satellite telecommunication networks such as Internet can provide the technical backbone to provide access to remote and widely distributed electronic databases to support the information needs of the health care community. Over the long term, access to such databases likely will be cost-effective, in contrast to the assuredly astronomical cost of building a comparable domestic print collection. The advent of new, low-cost electronic technologies probably will revolutionize health care information services in developing nations. However, for the immediate future, the medical libraries of Vietnam will require ongoing sustained support from the international community, so that minimal levels of resources will be available to support the information needs of the health care community. It is remarkable, and a credit to the determination of Vietnam's librarians that, in a country with a legacy of war, economic deprivation, and international isolation, they have somehow managed to provide a sound basic level of information services for health care professionals.

Communism↗
PubMed2004

[Construction of phage antibody library for Fab fragment from a convalescent patient infected with SARS coronavirus].

AIM: To construct the phage antibody library for Fab fragment from a convalescent patient infected with SARS coronavirus. METHODS: Total RNA was extracted from peripheral blood lymphocytes of a patient with SARS virus-specific IgG antibody and was transcribed reversely into cDNA. Using the cDNA as template, Fd and light chain of Fab genes were amplified by PCR with specific primer and were cloned into phagemid pComb3. Then the recombinant phagemid was electroporated into E.coli XL-1 Blue. The recombinant rate was detected by enzyme digestion analysis, and library repertoire was determined. RESULTS: The antibody library of Fab fragment from a convalescent patient infected with SARS conronavirus was constructed. The recombinant rate of light chain and heavy chain of Fd gene were 91% and 75%, respectively. The library size was 7.23 x 10(7). CONCLUSION: Phage antibody library for human Fab fragment has been constructed successfully, which lays the foundation for further study.

Antibodies, Viral↗
PubMed2004

[Construction of the subtracted cDNA libraries related to artemisinin-resistance of Plasmodium berghei].

OBJECTIVE: To construct the subtracted cDNA libraries related to artemisinin-resistance of Plasmodium berghei using suppression subtractive hybridization PCR (SSH PCR). METHODS: Total RNA was extracted from the artemisinin-sensitive (NS) and artemisinin-resistant (AR) strains of Plasmodium berghei K173. The cDNA synthesis followed the protocol of super SMART cDNA synthesis kit. Taking the NS as driver, AR as tester and reverse, two subtractions were performed by SSH PCR. Enriched different expressed cDNA was cloned into pMD18-T vector to construct subtractive libraries. RESULTS: The subtracted cDNA libraries of NS-AR and AR-NS contained 395 and 506 positive clones respectively. The PCR results of 108 clones picked randomly from each library showed 100 and 104 positive inserts contained in the plasmids respectively, and distributing in 250-2000 bp. CONCLUSION: The successful construction of the subtracted cDNA libraries related to artemisinin-resistance of P. berghei enable us to identify the different expressed genes involved in the resistance mechanism.

Animals↗
PubMed2003

[Construction and screening of TsF cDNA library specific for AchR].

OBJECTIVE: To construct the T suppress factor (TsF) cDNA library specific for AchR and screen out the positive clones of TsF. METHODS: The full-length cDNA was synthesized by the reverse transcripting from the TsF PolyA+mRNA, which had been extracted from the AchR specific suppressor lines (ARSL). The cDNA library was obtained using Lambda gt11 vector. Finally, this cDNA library was screened by Western blotting using Analysis of this cDNA library showed that titer of the monoclonal antibody specific for TCR-achain. RESULTS: recombinant bacteriophage was 1.4 x 10(7) pfu/ml, the rate of positive recombinant was 86.9%. The recombinant bacteriophage DNA were digested by EcoRI and electrophoresis analysis found that the main size of insertion was 0.8 to approximately 4.0 kb . As a result of screening, 27 positive clones were obtained. The positive recombinant bacteriophage DNA were digested by EcoRI, and electrophoresis analysis found that the size of insertion was 1 kb. Conclusion The results suggested that this cDNA library is available, the recombination phages can express TsF specific for AchR effectively and could be used to prepare the products of gene engineering of TsF.

Animals↗