摘要
采用逆转录-聚合酶链式反应(RT -PCR)方法,从小鼠黑色素瘤总RNA中扩增得到小鼠酪氨酸酶的cDNA .该基因重组到T7启动子控制下的表达载体pET - 2 2b(+)中,构建表达质粒pET -TYR并转化大肠杆菌Rosetta (DE3) .SDS -PAGE及蛋白质序列测定表明,经0 .8mmol/L异丙基硫代- β-D -半乳糖苷(IPTG)诱导后,可高效表达重组的小鼠酪氨酸酶,表达量约占菌体总蛋白质的2 5 % 。
Murine tyrosinase cDNA was amplified from total RNA of the murine melanotic carcinoma by reverse transcriptase PCR(RT-PCR). The expression plasmid pET-TYR was constructed by genetically cloning the DNA fragment into expression vector pET-22b(+) which under the control of T7 promotor. The results of SDS-PAGE and protein sequencing showed that after induction by 0.8?mmol/L IPTG, recombinant murine tyrosinase was highly expressed in E coli Rosetta(DE3). The quantities of expression product was up to 25% of the total protein of the bacteria in insoluble bodies form.
出处
《福州大学学报（自然科学版）》
CAS
CSCD
北大核心
2005年第2期254-258,共5页
Journal of Fuzhou University(Natural Science Edition)
基金
福建省自然科学基金重大科技项目 (2 0 0 1F0 0 9)
福州大学发展基金资助项目 (2 0 0 2 -XY - 11
XJY - 0 2 4 1
XKJ(QD) - 0 132 )
