猪乳铁蛋白N-叶基因PLF-N在大肠杆菌中的克隆、表达及特性分析  

Cloning,Expression and Characteristic Analysis of N-lobe PLF-N Gene of Porcine Lactoferrin in Escheichia coli

作　　者：汪以真[1] 刘光富[1] 初晓娜[1] 黄海青[1] 

机构地区：[1]浙江大学饲料科学研究所

出　　处：《农业生物技术学报》2005年第5期635-638,共4页Journal of Agricultural Biotechnology

基　　金：国家教育部新世纪优秀人才(No.NCET040543);浙江省重点科研项目(No.2005C22048)资助

摘　　要：从泌乳15d的约克夏母猪乳腺组织中提取总RN A,用RT-PCR技术扩增PLF-N基因,获得1条1038bp的目的片段。将目的基因克隆到质粒载体pET-28a(+)中,并转化大肠杆菌(Escherichia coli)感受态细胞BL21(D E3),经IPTG诱导后进行SD S-PA G E和W estern-blot分析。结果表明,PLF-N基因在大肠杆菌BL21中表达,表达产物分子量约为42kD。To study the relationship between the structure and function of porcine lactoferrin N-lobe, the total RNA in mammary gland cells of 15-day lactating York Shine sow was extracted and PLF-N gene was amplified using RT-PCR. A DNA fragment about 1 038 bp in length was obtained and the PCR product was cloned into Escheichia coil expression vector pET-28a（＋）. The recombinant plasmid pET-PLF-N then was transformed to competent cell BL21, and expressed at 37℃ by IPTG induction. Recombinant protein was analyzed with SDS-PAGE and Western blot. Both results showed that the rPLF-N was well expressed in E.coli and molecular weight of the recombinant product was estimated to be approximately 42 kD.

关 键 词：猪乳铁蛋白N-叶 基因 克隆 表达 

分 类 号：Q78[生物学—分子生物学] Q785