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The Primary Study of the Distribution of the EnJSRV in the Genome of Mongolian Sheep

Author: ChaiJu
Tutor: ZhangWenGuang;LiuShuYing;LiJinQuan
School: Inner Mongolia Agricultural University
Course: Animal Genetic Breeding and Reproduction
Keywords: Ovine pulmonary adenocarcinoma(OPA) Jaagsiekte sheep retrovirus(JSRV) Endogenous Jaagsiekte sheep retrovirus(enJSRV) Clone PCR FISH
CLC: S826
Type: Master's thesis
Year: 2008
Downloads: 28
Quote: 2
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In order to study the distribution of the endogenous pulmonary adenomatosis virus gene in the chromogenes of the Mongolia sheep, 5 pairs of primers were designed according to the whole sequence of the enJSRV(DQ838493)issued by Inner Mongolia Agriculture University in GenBank, the primers of the wings of the exogenous pulmonary adenomatosis virus gene inserting site in the pulmonary SP-A gene and the primer of the long terminal repeat of the enJSRV6 and enJSRV10. The total DNA was extracted from the blood from the jugular of the healthy sheep. The SP-A protein gene and the LTR of the enJSRV6 and enJSRV10 were amplified through PCR with the total DNA as the template which resulted in 5 specific gene fragments which were further cloned into the pMD19-T vector and sequenced. The resulting products were duplexing sequenced and then spliced and analyzed with the DNAstar software. The results showed that there was no enJSRV gene in the SP-A gene where the former could insert, but there was the insertion site for the enJSRV6 and enJSRV10 in the genome of the Mongolian sheep. The analysis of the ends’LTR of the enJSRV6 and enJSRV10 showed the typical structure of the LTR in the endogenous pulmonary adenomatosis gene of the Mongolian sheep and the stabilization of the ends’U3(R)U5 in each endogenous virus. Only the U3 was the variable region, and the R and U5 were rather conservative. To further study the distribution of the genes of the enJSRV in the genome of the Mongolian sheep, 1 pair of the primer was designed according to the gag gene in the enJSRV(DQ838493)genome, and the probes were synthesized with the purified PCR products and the FISH was performed. The hybridization specimen made of the metaphase chromosome from the peripheral blood lymphocyte of the sheep, through the karyotype analysis( the length and the chromosome located), were compared with the sheep standard G-band ideograph, and the gag gene was then located. And the location and distribution of the gene were then determined primarily, there were the gag gene in all the 6 chromosomes, 1q45, 1p23, 2q41, 3q23, 6q13, 9q22 and 14q25of the Mongolian sheep.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Sheep
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