Rubisco小亚基基因片段的亚克隆及其瞬时表达载体的构建  

Subcloning of Rubisco Small Subunit Gene Fragment and the Construction of Transient Expression Vector

作　　者：麻鹏达[1] 周晓馥[2] 秦波[1] 董春光[1] 王兴智[1] 

机构地区：[1]东北师范大学遗传与细胞研究所,吉林长春130024 [2]吉林师范大学基因工程研究所,吉林四平136000

出　　处：《分子植物育种》2004年第4期527-530,共4页Molecular Plant Breeding

基　　金：国家自然科学基金资助(30370767)。

摘　　要：1,5-二磷酸核酮糖羧化酶/加氧酶位于植物叶绿体中,是绿色植物中含量最丰富的蛋白,它催化CO2固定和光呼吸作用的第一步。从含Rubisco小亚基基因片段的pGR107-rbcS质粒中亚克隆出Rubisco小亚基基因片段,将PCR产物与pGEM-T Easy载体连接,用EcoRI酶切鉴定酶切重组载体,利用低溶点胶回收大约300bp大小的Rubisco小亚基基因片段,将其回收片段与pSLJ75515瞬时表达载体连接,用EcoRI酶切鉴定,筛选出阳性重组体,即为pSLJ75515-rbcS瞬时表达载体。本实验为进一步研究Rubisco小亚基基因功能和基因沉默机制奠定了基础。Ribusco-1,5-bisphosphate (RuBP) carboxylase/oxygenase (Rubisco) located in the chloroplast is the most abundant protein in the leaves of light-grown plants. The enzyme catalyzes the first step in CO2 fixation and photorespiration. Rubisco small subunit gene fragment was subcloned in pGEM-T Easy vector from plasmid pGR107-rbcS containing Rubisco small subunit gene fragment. After verification by restriction analysis, pSLJ75515-rbcS transient expression vector containing about 300 bp Rubisco small subunit gene fragment was successfully constructed by means of recombinant DNA technology. These results could be used for further research of Rubisco subunit gene function and mechanism of RNA silencing.

关 键 词：Rubisco小亚基基因 基因片段 亚克隆 瞬时表达载体 基因构建 植物 叶绿体 

分 类 号：S336[农业科学—作物遗传育种]