期刊论文

中文

华中科技大学学报(医学版)

1672-0741

2004

33

3

265-268,272

10.3870/j.issn.1672-0741.2004.03.006

国家自然科学基金资助项目（No.39970307）;国家自然科学基金资助项目（No.39970307）

本校为其他机构

目的 获得人源受体相关蛋白(RAP)，对表达条件进行优化。方法 RAP cDNA被插入到原核表达质粒 pT7-L和pET-28a(+)中，分别构建成RAP重组表达载体，转化两种BL21菌株后，转化菌用异丙基硫代半乳糖苷(IPTG)诱导，选择pT7-PL-RAP质粒进行表达条件的优化，采用镍离子亲和层析纯化所获得的RAP，通过检测与巨噬细胞的结合能力评估其生物活性。结果 两个重组质粒均获得了RAP的高表达，对于pT7-PL-RAP质粒， RAP在BL21(DE3，plysS)菌株中的表达显著高于在BL21(DE3)菌株的表达；有氯霉素存在时表达高于无氯霉素时；诱导表达的RAP大部分为可溶性的；镍离子亲和层析树脂纯化所获得的RAP，可与富含低密度脂蛋...

Objective To obtain human receptor associated protein and to optimize its expression con-ditions. Methods RAP cDNA was inserted into prokaryotic expression vector-pT7-PL and pET-28a ( + ) to construct two RAP recombinant human RAP expression plasmids. After they were transformed to the BL21 strains, the transformed strains were induced with isopropylthiogalactoside (IPTG). The pT7-PL-RAP was selected for optimal expression, and Ni' -nitrilotriacetic acid (Ni-NTA) affinity chromatogram was applied for purification of RAP. The biological activity of RAP was assessed by detecting its binding abil...