Cloning,Expression of Ribosome Inactivating Proteins Gene from Atriplex patens in Xinjiang
SUN Su-Rong, HU Jun, LAN Hai-Yan, ZHANG Fu-Chun*
(Xingjiang Key Laboratory of Biological Resourses and Genetic Engineering,College of Life Science and Technology,Xinjiang University,Urumqi 830046,China)
Abstract：The complete open reading frame (ORF) for the ribosome-inactivating proteins (RIPs) in the Atriplex patens leaves was obtained by RT—PCR．The sequence data showed that the cloned fragment is 843bp. The open reading frame of the fragment contains 281 amino acid residues and there is a signal sequence with 26 amino acid residues at its N-terminus. The analysis suggests that ApRIP belongs to type ⅠRIP. The amplified fragment has 82%，55% and 41% identity comparing with RIP gene nucleotide of Chenopodium album, pokeweed antiviral protein and Trichosanthes kirilowii trichosanthin, respectively.The amino acid sequence and SWISS-MODEL analysis indicated that the “EAARXKXY”is active centre of the enzyme in RIPs. The IPTG inducible expression vector containing the RIP gene was constructed and transferred into E coli BL21(DE3).The specific protein was produced after inducing with 0．4mmol／L IPTG SDS—PAGE of the fusion protein demonstrated that the RIP migrated at a size of 77kDa．The immunization was performed by intra—muscular injection of pCDNA3-Aprip，and then antiserum was detected by ELISA．Western blotting analysis showed the antiserum was specific against the ribosome inactivating proteins．
孙素荣,胡俊,兰海燕,张富春. 滨藜（Atriplex patens）核糖体失活蛋白基因的克隆和分析[J]. 中国生物化学与分子生物学报, 2008, 24(03): 281-285.
SUNSu-Rong,HUJun,LANHai-Yan,ZHANGFu-Chun. Cloning,Expression of Ribosome Inactivating Proteins Gene from Atriplex patens in Xinjiang. Chinese Journal of Biochemistry and Molecular Biol, 2008, 24(03): 281-285.
