Construction of Genomic DNA Libraries in Cosmid Vectors

2006 ◽  
Vol 2006 (1) ◽  
pp. pdb.prot4000
Author(s):  
Joseph Sambrook ◽  
David W. Russell
Keyword(s):  
Genomic Dna ◽  
Dna Libraries

scholarly journals Absence of Replication-Competent Human-Tropic Porcine Endogenous Retroviruses in the Germ Line DNA of Inbred Miniature Swine

2004 ◽  
Vol 78 (5) ◽  
pp. 2502-2509 ◽  
Author(s):  
Linda Scobie ◽  
Samantha Taylor ◽  
James C. Wood ◽  
Kristen M. Suling ◽  
Gary Quinn ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Germ Line ◽  
Human Cells ◽  
Endogenous Retroviruses ◽  
Open Reading Frames ◽  
Miniature Swine ◽  
Dna Libraries ◽  
Porcine Endogenous Retroviruses ◽  
Perv Transmission

ABSTRACT The potential transmission of porcine endogenous retroviruses (PERVs) has raised concern in the development of porcine xenotransplantation products. Our previous studies have resulted in the identification of animals within a research herd of inbred miniature swine that lack the capacity to transmit PERV to human cells in vitro. In contrast, other animals were capable of PERV transmission. The PERVs that were transmitted to human cells are recombinants between PERV-A and PERV-C in the post-VRA region of the envelope (B. A. Oldmixon, J. C. Wood, T. A. Ericsson, C. A. Wilson, M. E. White-Scharf, G. Andersson, J. L. Greenstein, H. J. Schuurman, and C. Patience, J. Virol. 76:3045-3048, 2002); these viruses we term PERV-A/C. This observation prompted us to determine whether these human-tropic replication-competent (HTRC) PERV-A/C recombinants were present in the genomic DNA of these miniature swine. Genomic DNA libraries were generated from one miniature swine that transmitted HTRC PERV as well as from one miniature swine that did not transmit HTRC PERV. HTRC PERV-A/C proviruses were not identified in the germ line DNAs of these pigs by using genomic mapping. Similarly, although PERV-A loci were identified in both libraries that possessed long env open reading frames, the Env proteins encoded by these loci were nonfunctional according to pseudotype assays. In the absence of a germ line source for HTRC PERV, further studies are warranted to assess the mechanisms by which HTRC PERV can be generated. Once identified, it may prove possible to generate animals with further reduced potential to produce HTRC PERV.

A simplified procedure for cDNA and genomic library construction using nonpalindromic oligonucleotide adaptors

1989 ◽  
Vol 67 (4-5) ◽  
pp. 205-209 ◽  
Author(s):  
Cynthia R. D'Souza ◽  
Ken V. Deugau ◽  
John H. Spencer
Keyword(s):  
Genomic Dna ◽  
Restriction Site ◽  
Genomic Library ◽  
Restriction Endonuclease Site ◽  
Dna Libraries ◽  
Target Dna ◽  
Restriction Sites ◽  
Enzyme Cleavage ◽  
Simplified Procedure ◽  
Genomic Cdna

The properties and characteristics of oligonucleotide adaptors for use in a simplified procedure for the construction of cDNA and genomic DNA libraries are described. The adaptors are suitable for joining to blunt ended cDNA or sheared genomic DNA, and then to the cohesive ends of restriction sites in vectors. Each adaptor consists of two oligonucleotides with complementary but nonpalindromic sequences that include an internal restriction site, a 5′ phosphorylated blunt end, and an overlapping or staggered 5′ hydroxylated end corresponding to a restriction endonuclease site in a vector of choice. Ligation of the blunt end to high molecular weight target DNA proceeds efficiently and there is no tandem concatenation of the adaptor. Insertion into the appropriate vector only requires ligation of the cohesive ends. There is no requirement for methylation, restriction enzyme cleavage, G-C tailing, or denaturation after ligation of the adaptor to the target DNA, all characteristics of other procedures.Key words: library, genomic, cDNA, oligonucleotides, adaptors.

Genomic DNA Libraries

1987 ◽  
Vol 00 (1) ◽  
Author(s):  
David D. Moore
Keyword(s):  
Genomic Dna ◽  
Dna Libraries

scholarly journals Universal Linker and Ligation Procedures for Construction of Genomic DNA Libraries Enriched for Microsatellites

BioTechniques ◽  
1999 ◽  
Vol 27 (3) ◽  
pp. 500-507 ◽  
Author(s):  
Matthew B. Hamilton ◽  
Elaine L. Pincus ◽  
Anthony Di Fiore ◽  
Robert C. Fleischer
Keyword(s):  
Genomic Dna ◽  
Dna Libraries

scholarly journals Genomic organization and in vivo characterization of proteolytic activity of FtsH of Mycobacterium smegmatis SN2

Microbiology ◽  
2004 ◽  
Vol 150 (8) ◽  
pp. 2629-2639 ◽  
Author(s):  
Gopalakrishnapillai Anilkumar ◽  
Ramanujam Srinivasan ◽  
Parthasarathi Ajitkumar
Keyword(s):  
Mycobacterium Smegmatis ◽  
Genomic Dna ◽  
Genomic Organization ◽  
Growth Arrest ◽  
Upstream Region ◽  
E Coli ◽  
Dna Libraries ◽  
In Vivo Characterization

The ftsH gene of Mycobacterium smegmatis SN2 (MsftsH) was cloned from two independent partial genomic DNA libraries and characterized, along with the identification of ephA and folE as the neighbouring upstream and downstream genes respectively. The genomic organization of the MsftsH locus was found to be identical to that of the Mycobacterium tuberculosis ftsH gene (MtftsH) and similar to that of other bacterial genera, but with divergence in the upstream region. The MsftsH gene is 2·3 kb in size and encodes the AAA (ATPases Associated with diverse cellular Activities) family Zn2+-metalloprotease FtsH (MsFtsH) of 85 kDa molecular mass. This was demonstrated from the expression of the full-length recombinant gene in Escherichia coli JM109 cells and from the identification of native MsFtsH in M. smegmatis SN2 cell lysates by Western blotting with anti-MtFtsH and anti-EcFtsH antibodies respectively. The recombinant and the native MsFtsH proteins were found localized to the membrane of E. coli and M. smegmatis cells respectively. Expression of MsFtsH protein in E. coli was toxic and resulted in growth arrest and filamentation of cells. The MsftsH gene did not complement lethality of a ΔftsH3 : : kan mutation in E. coli, but when expressed in E. coli cells, it efficiently degraded conventional FtsH substrates, namely σ 32 protein and the protein translocase subunit SecY, of E. coli cells.

CONSTRUCTION AND SCREENING OF cDNA AND GENOMIC DNA LIBRARIES FOR HI STONE CODING SEQUENCES FROM MOUSE EMBRYOS

1981 ◽  
Vol 9 (2) ◽  
pp. 256P-256P
Author(s):  
J. H. McVey ◽  
R. Krumlauf ◽  
S. Graham ◽  
M. L. M. Anderson ◽  
C. McAllister
Keyword(s):  
Genomic Dna ◽  
Mouse Embryos ◽  
Coding Sequences ◽  
Dna Libraries

scholarly journals Construction of tomato genomic DNA libraries in a binary-BAC (BIBAC) vector

1999 ◽  
Vol 18 (2) ◽  
pp. 223-229 ◽  
Author(s):  
Carol M.. Hamilton ◽  
Anne Frary ◽  
Yimin Xu ◽  
Steven D.. Tanksley ◽  
Hong-Bin Zhang
Keyword(s):  
Genomic Dna ◽  
Dna Libraries

scholarly journals Construction of representative genomic DNA libraries using one microgram of DNA

1989 ◽  
Vol 17 (2) ◽  
pp. 818-818 ◽  
Author(s):  
Ming Lu
Keyword(s):  
Genomic Dna ◽  
Dna Libraries
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