Cloning and Identifying Expression Vectors of α-1,3 Galactosyltransferase Gene from Bama Minipig in Guangxi

doi:10.3969/j.issn.1674-5817.2012.01.001

Abstract

Abstract: Objective For the purpose of producing GGTA1 gene silencing bama-mini pig, cloned and analyzed the function of α-1,3 galactosyltransferase gene of bama-mini pig then constructed two expression vectors. Methods The GGTA1 gene was amplified from liver tissue by RT-PCR using a specific pair of primers which were designed and synthesized according to the GGTA1gene sequence released in GenBank, the amplified cDNA fragment was ligated into pMD18-T vector after purification. The recombinant was verified by the method of PCR and sequencing, then the enzyme digested products was ligated into pEGFP-N₁ to construct the expression vectors. identified by PCR and digesting reaction. Results The α-1,3 galactosyltransferase gene of bama-mini pig exist the absence of 5-exon(81-116) which containing 1080 bp, and existence of 5-exon which containing 1116 bp. Conclusions There exit the absence of 5-exon in the bama-mini pig, Construction of the both expression vectors (pEGFP-GGTA1-1 and pEGFP-GGTA1-2) can be transfected into PK-15 cells to identify their functional expression, also can be used in the experiment of gene silencing pig α-1,3 galactosyltransferase gene.

Key words: Bama-minipig in Guangxi, α-1, 3galactosyltransferase gene, Eukaryotic expression vector, Expression identification

CLC Number:

R332
Q95-33

GUO Xiao-ping, CHEN Shi-jin, LAN Gan-qiu, JIANG Qin-yang, GUO Ya-fen. Cloning and Identifying Expression Vectors of α-1,3 Galactosyltransferase Gene from Bama Minipig in Guangxi[J]. Laboratory Animal and Comparative Medicine, 2012, 32(1): 1-7.

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