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Cloning and characterization of goat enteric _Ò-defensin cDNA.
2010
Kumar, Ashish | Sharma, Ambika | Kumar, Ashok | Dev, Kranti
The mRNA from distal ileum of Indian goat was cloned and characterized after purification. cDNA was synthesized using goat ileal epithelial RNA, omniscript and sensiscript reverse transcriptase and amplified by Hotstart Taq DNA polymerase with primers designed by taking conserved regions of cattle enteric _Ò-defensin, cattle lingual antimicrobial peptide(LAP) and goat _Ò-defensin-2 sequences. The amplified cDNA of 253bp was, ligated to linearised TA cloning vector and transformed into XLblue strain of E.coli which was grown overnight at 37oC in a LB plate containing ampicillin, IPTG and X-Gal. The recombinant plasmid was isolated and digested with NcoI. The white colonies showed a release of 253bp insert. The sequence analysis showed 26, 16 and 5-nucleotide substitution having 85.6%, 91.3%, 97.4% homology with reported cattle EBD, buffalo EBD and goat BD2 mRNA respectively. The deduced amino acid sequence encodes for a 64 amino acid precursor peptide showing 12,18 and 4 amino acid substitution having 80%, 70.8%, 93.8% homology with buffalo EBD, cattle EBD and goat BD2 peptide respectively. Both nucleotide and amino acid sequence homology showed that the cloned sequence was closer to goat BD2.
Show more [+] Less [-]Cowdria ruminantium DNA is unstable in a SuperCos 1 library
1999
Brayton, K.A. | De Villiers, E.P. | Fehrsen, J. | Nxomani, C. | Collins, N.E. | Allsopp, B.A. (Agricultural Research Council, Onderstepoort (South Africa). Onderstepoort Veterinary Inst.)
Production of cloning animals by fresh and frozen-thawed nuclear transfer embryos 2
1993
Hwang, W.S. | Cho, C.H. | Lee, C.W. | Lee, B.C. (Seoul National University, Suwon (Korea Republic). College of Veterinary Medicine)
Cloning and sequencing of the gene encoding structural protein of canine parvovirus isolated in Korea
1992
Park, J.H. | Song, J.Y. | Lee, J.B. | Hyun, B.H. | An, S.H. (Rural Development Administration, Anyang (Korea Republic). Veterinary Research Inst.) | Jun, M.H. (Chungnam Nat'l Univ., Taejon (Korea Republic). Coll. of Veterinary Medicine)
New trends of vaccine development: Recombinant vaccinia viruses (expression vectors) as vaccines
1989
Kim, U.H. (Kangwon National Univ., Chunchon (Korea R.). Coll. of Animal Agriculture)
The prospect of live vaccines consisting of genetically modified vaccinia virus expressing foreign genes is exciting, but important issues concerning safety and efficacy need to resolved. Vaccinia virus (VV) is an efficient expression vector with broad host range infectivity and large DNA capacity. This vector has been particularly useful for identifying target antigens for humoral and cell-mediated immunity. The WHO smallpox eradication program, involving the extensive use of VV vaccines, resulted in the late 1970s in the elimination of one of the world's most feared diseases. This achievement is a triumph for preventive medicine and for international collaboration in public health. In 1980, WHO recommended that the routine use of smallpox vaccine should be stopped. Against this background, the prospect of live vaccines consisting of genetically modified VV expressing foreign antigens arising from the work of Moss, and Paoletti and their colleagues in 1982 has been greeted with enthusiasm. These investigators have shown that genes coding for immunogenic proteins can be inserted into VV DNA without impairing the ability of the virus to grow in cell culture. Moreover experimental animals infected with VV recombinants containing genes coding for a variety of immunizing proteins have been shown to be protected against challenge infection with the corresponding infectious agent. In this communication, I describe current progress in the construction of a novel plasmid vector that facilitates the insertion and expression of foreign genes in VV as well as the selection of recombinants
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