Biotechnology Bulletin ›› 2015, Vol. 31 ›› Issue (7): 109-114.doi: 10.13560/j.cnki.biotech.bull.1985.2015.07.016

• Research report • Previous Articles     Next Articles

Construction and Application of the RNAi Vector for BBS1 Gene in Chlamydomonas reinhardtii

Chen Lihong, Li Lili, Gao Lifen, Zhou Junfei, Li Tiantian, Peng Hai   

  1. (Institute for System Biology,Jianghan University,Wuhan 430056)
  • Received:2014-11-12 Online:2015-07-16 Published:2015-07-16

Abstract:

Bardet-Biedl syndrome is a disease caused by mutations of various genes and associated with afunction of primary cilia. There are 12 disease-causing genes of BBS1-12 respectively. This study aimed to understand the function of BBS1 gene in Chlamydomonas reinhardtii through RNAi interference technology. Firstly, the fast and simple RNAi backbone vector was constructed. Subsequently, a cDNA in BBS1 gene was cloned into media vector pEASY-T1 using RT-PCR. After sequencing, the target fragment was inserted into the RNAi backbone vector through the twice enzyme digestion. Verified by PCR testing, enzyme digestion and sequencing, the RNAi vector of BBS1 gene was successfully constructed. The phenotype of transgenic C. reinhardtii CC503 transformed into BBS1 RNAi plasmid by gene gun was significantly different from that of the control and their phototaxis changed compared to the control.

Key words: Chlamydomonas reinhardtii, RNAi vector, BBS1 gene, phototaxis