猪血清白蛋白基因的克隆与序列分析

Cloning and Sequence Analysis of Porcine Serum Albumin Gene

血清白蛋白融合技术是一种开发长效重组蛋白药物的新技术。为了获得猪血清白蛋白(PSA)基因,根据GenBank(登录号:AY663543)中PSA的基因序列,设计一对引物,采用RT-PCR从新鲜猪肝组织中扩增出PSA基因的全长cDNA,产物纯化后连接至pBS-T载体,转化大肠埃希菌TG1,采用PCR法及酶切鉴定法筛选出阳性菌株后进行测序。序列分析结果表明,成功克隆了PSA基因的1 821 bp全长cDNA,与GenBank中参照基因序列同源性为99.6%,氨基酸序列同源性为99.5%。本研究数据已提交至GenBank,并申请到一个登录号EF202601。

Albumin fusion technology was a novel technology utilized to develop long-acting biologics. According to the cDNA sequence of PSA （porcine serum albumin） derived from GenBank （Accession No. AY663543）, a pair of primers were designed. Porcine RNA was isolated from porcine liver cells and the cDNA coding PSA was amplified by RT-PCR subsequently. The DNA fragment was cloned into vector pBS-T and transformed into E. coli TG1. After identified by both PCR and restrictive endonuclease digestion, the positive recombinant plasmid was sent for sequencing. The sequencing result suggested that the coding PSA gene was composed of 1 821 bp. The nucleotide sequences and the deduced amino sequences of PSA-T were compared with PSA. The nucleotide homology between them was 99.6 %, the amino acid homology was 99.5%. The research data had been submitted to GenBank and gained a new Accession No. EF202601.