水生生物学报2012，Vol.36Issue(4)：640-645,6.DOI:10.3724/SP.J.1035.2012.00640
雨生红球藻质体球滴结构蛋白基因的克隆与原核表达
CLONING AND PROKARYOTIC EXPRESSION OF THE PLASTOGLOBULES PROTEIN GENE FROM HAEMATOCOCCUS PLUVIALIS
摘要
Abstract
Carotenoids were organized in plastoglobules located in the interthylakoid space of the chloroplast for many plants. However, the plastoglobule, a carotenoid deposit structure in chromoplast, was not clear in Haematococcus pluvialis yet up to date. In the present study, many osmiophilic globuli appeared in both the chromoplast and cytoplasm ofH, pluvialis under electron microscopy with a modified method. Those revealed clearly the presence of plastoglobules in the chloroplast of H. pluvialis. A full-length cDNA was isolated from H. pluvialis through a RT-PCR and RACE (rapid amplification of cDNA ends) method. And the completing cDNA encoded a protein which had high homology with the major component proteins of plastoglobules in many plants and was termed as HPGP (Haematococcus plastoglobules protein). Furthermore, this cDNA was expressed in the prokaryotic expression system, and the resultant fusion proteins with His-Tags were purified by immobilized metal affinity chromatography (IMAC). The polyclonal antibodies against the protein were raised in rabbits and the raw serum was collected without further purification. Those might provide an important basis for unraveling the function of the protein in the future.关键词
雨生红球藻/虾青素/质体球滴/基因克隆/原核表达/抗体Key words
Haematococcus pluvialis/ Astaxanthin/ Plastoglobules/ Gene cloning/ Prokaryotic expression/ Antibody分类
生物科学引用本文复制引用
范勇,于广欣,汪乐霓,曲芳兵,冷鹂,兰利琼,卿人韦..雨生红球藻质体球滴结构蛋白基因的克隆与原核表达[J].水生生物学报,2012,36(4):640-645,6.基金项目
国家高技术发展计划(863计划)(2007AA09Z427) （863计划）
国家自然科学基金面上项目(40976092) （40976092）
四川省科技厅应用基础项目(2008JY0030)资助 （2008JY0030）
