The porcine prothymosin a (ProTα) cDNA fragment was amplified from kidney of por-cine by reverse transcription-polymerase chain reaction (RT-PCR). Sequence analysis showed that the complete open reading frame (ORF) of ProTα gene includes 333 bp and encodes 109 aa. The sequence shared 99.4% homology with the two published ProTα genes, pET32-mProTα was constructed by gene rearrangement, then it was transformed into E. coli Rosetta 2 (DE3). SDS-PAGE electrophoresis analysis showed that the fusion protein of ProTα molecular mass was about 12 070 of molecular weight. MTT essay confirmed that it can enhance lymphocyte proliferation obviously.%应用RT-PCR技术从猪肾脏中扩增到猪前胸腺素基因,测序结果表明,猪前胸腺素完整开放阅读框(ORF)共333 bp,编码109 aa,与已公布的2个猪前胸腺素基因核苷酸同源性均为99.4%.构建了重组质粒pET-32-mProTα,并转化大肠杆菌(E.coli)Rosetta 2(DE3).SDS-PAGE电泳和免疫印迹分析显示,表达产物约占菌体总蛋白的40%,相对分子质量约为12.07 ku.MTT法试验证实,表达产物初步纯化、透析复性后,可明显增强淋巴细胞的增殖.
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