摘要
应用反转录-聚合酶链式反应(RT PCR)技术,通过一对自行设计的引物,从经ConA诱导活化的鸡脾淋巴细胞的RNA中扩增出一特异性基因片段,其大小约为500bp。该片段经酶切和测序鉴定为鸡IFN γ基因。将其插入原核表达载体pGEX 4T 1,并导入大肠杆菌BL21细胞中,经IPTG诱导培养,获得分子量约为43000的蛋白带,表明所克隆的CHIFN γ基因在原核细胞中得到表达。
A specific DNA fragment with a length of 500 bp was cloned from concanavalin A (ConA)-activated spleen lymphocytes by reverse transcription-polymerase chain reaction (RT-PCR) with a pair of designed primers. By means of endonuclease and sequencing, this fragment was identified as the gene of chicken interferon-gamma (CHIFN-γ),it was inserted into PGEX-4T-1, a prokaryotic expression plasmid, and expressed in Escherichia coli BL21, which were further induced by IPTG and cultured,and a fusion protein was obtained with about 43 000 of molecular weight in SDS-PAGE. This result showed that the cloned CHIFN-γ gene was expressed in prokaryotic cells.
出处
《安徽农业大学学报》
CAS
CSCD
北大核心
2004年第1期92-95,共4页
Journal of Anhui Agricultural University
基金
国家自然基金项目(30270974)
安徽省"十五"科技攻关项目(01013003)资助。
