%0 Journal Article
%T 菠菜43 kD叶绿素？？a？？结合蛋白编码基因克隆及原核表达条件优化<br>Cloning and prokaryotic expression of spinach 43 kDa chlorophyll a binding protein gene in ？？Escherichia coli
%A 刘骥
%A 王豪
%A 肖清洁
%A 谢思思
%A 杜林方
%J 四川大学学报 (自然科学版)
%D 2015
%X 将菠菜43 kD叶绿素结合蛋白编码基因亚克隆至原核表达载体pET 28a上, 构建重组表达质粒pET 28a ？？psbC？？转化感受态？？E.coli ？？BL21 (DE3), 经IPTG诱导实现了外源基因的可溶性表达. 探讨了含组氨酸标签融合蛋白的最佳表达条件（包括表达单克隆、时间、温度对表达的影响）. 利用Ni？？？？2+？？ Sepharose 4 Fast Flow亲合层析纯化出His apo CP43蛋白. 体外重组实验证实His apo CP43能特异结合叶绿素？？a？？, 经过温和电泳分离纯化所得体外重组色素蛋白复合物具有与提取自类囊体膜的天然CP43相似（但不完全相同）的荧光特性.<br>The ？？psbC？？ gene isolated from Spinach was expressed in ？？Escherichia coli？？ in soluble state. The recombinant expression plasmid pET 28a ？？psb？？C containing spinach apo CP43 gene was transformed into competent cell of ？？E.coli？？ BL21 (DE3). The positive colony expressed effectively His tag containing soluble apo CP43 fusion protein in ？？E.coli？？ BL21 (DE3) when induced by IPTG under the optimal condition. The heterogeneous expression condition, including the expression monocolony, time, IPTG and temperature, was explored further. Near homogeneous purified His apo CP43 fusion protein judged by SDS PAGE was obtained by Ni Sepharose Affinity Chromatography. ？？In vitro？？ reconstitution experiment was carried out and the formation of stable chlorophyll ？？a？？ protein complex was analyzed by partially denaturing polyacrylamide green gel electrophoresis. After electrophoresis, the gel band containing blue green colour was excised and measured by fluorescence emission and excitation spectra. The overall spectrum is (in a first approximation) similar to native CP43
%K 43 kD叶绿素？？a？？结合蛋白 原核表达 条件优化 色素重组 荧光性质？？&lt
%K br&gt
%K His apo CP43 fusion protein
%K prokaryotic expression
%K purification
%K reconstitution experiment
%K fluorescence
%U http://science.ijournals.cn/jsunature_cn/ch/reader/view_abstract.aspx?file_no=20151035&flag=1