Recombination and Heterologous Expression of Aliophycocyanin Gene in the Chloroplast of Chlamydomonas reinhardtii  

作　　者：Zhong-Liang SU Kai-Xian QIAN Cong-Ping TAN Chun-Xiao MENG Song QIN 

机构地区：[1]Department of Biotechnology, College of Life Sciences, Zhejiang University Hangzhou 310027, China [2]Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China [3]Qingdao University of Science and Technology, Qingdao 266061, China [4]Graduate University of Chinese Academy of Sciences, Beijing 100039, China

出　　处：《Acta Biochimica et Biophysica Sinica》2005年第10期709-712, ,共4页生物化学与生物物理学报（英文版）

基　　金：This work was supported by a grant from the Key Innovative Project (KZCX-3-SW-215)of the Chinese Academy of Sciences

摘　　要：Heterogeneous expression of multiple genes in the nucleus of transgenic plants requires the introduction of an individual gene and the subsequent backcross to reconstitute multi-subunit proteins or metabolic pathways. In order to accomplish the expression of multiple genes in a single transformation event, we inserted both large and small subunits of allophycocyanin gene （apcA and apcB） into Chlamydomonas reinhardtii chloroplast expression vector, resulting in papc-S. The constructed vector was then introduced into the chloroplast of C. reinhardtii by micro-particle bombardment. Polymerase chain reaction and Southern blot analysis revealed that the two genes had integrated into the chloroplast genome. Western blot and enzyme-linked immunosorbent assay showed that the two genes from the prokaryotic cyanobacteria could be correctly expressed in the chloroplasts of C. reinhardtii. The expressed foreign protein in transformants accounted for about 2%-3% of total soluble proteins. These findings pave the way to the reconstitution of multi-subunit proteins or metabolic pathways in transgenic C. reinhardtii chloroplasts in a single transformation event.Heterogeneous expression of multiple genes in the nucleus of transgenic plants requires the introduction of an individual gene and the subsequent backcross to reconstitute multi-subunit proteins or metabolic pathways. In order to accomplish the expression of multiple genes in a single transformation event, we inserted both large and small subunits of allophycocyanin gene （apcA and apcB） into Chlamydomonas reinhardtii chloroplast expression vector, resulting in papc-S. The constructed vector was then introduced into the chloroplast of C. reinhardtii by micro-particle bombardment. Polymerase chain reaction and Southern blot analysis revealed that the two genes had integrated into the chloroplast genome. Western blot and enzyme-linked immunosorbent assay showed that the two genes from the prokaryotic cyanobacteria could be correctly expressed in the chloroplasts of C. reinhardtii. The expressed foreign protein in transformants accounted for about 2%-3% of total soluble proteins. These findings pave the way to the reconstitution of multi-subunit proteins or metabolic pathways in transgenic C. reinhardtii chloroplasts in a single transformation event.

关 键 词：Chlamydomonas reinhardtii chloroplast transformation allophycocyanin gene 

分 类 号：Q949.2[生物学—植物学]