Alternative splicing and expression of the Alboserpin-1 from salivary gland of Aedes albopictus
CHENG Jin-zhi1, 3, SUN Yu2, CHEN Lu2, LIU Jian1, WU Jia-hong1
(1.Department of Parasitology, Guiyang Medical College, Guiyang 550004, China; 2.The Affliated Hospital of Guiyang Medical College, Guiyang 550004, China; 3.Laboratory for Modern Pathogen Biology, Guiyang 550004, China)
Abstract：To clone and prokaryotically express the Aalbserpin-1 gene from the salivary gland of Aedes albopictus, the full-length cDNA sequence of Aalbserpin-1 gene was amplified by RT-PCR, then constructed into the prokaryotic expression vector pET28a(+) and expressed in E. coli BL21(DE3) with IPTG induction. The recombinant protein was detected by SDS-PAGE and purified by Ni-IDA affinity chromatography. An expression analysis was conducted by real-time RT-PCR. It was demonstrated that two alternative transcripts were obtained (Aalbserpin-1_1 and Aalbserpin-1_2). To compare with Aalbserpin-1 (AY826096.1), Aalbserpin1_1 and Aalbserpin-1_2 shared 95% and 90% similarity respectively. An alternative splicing exon (24 amino acids: NLLTNRIVSQSSYRRVAIYDFISG) in Aalbserpin1_2 was just located in the reactive centre loop (RCL) of SERPIN, a primary determinant of functionality. A recombinant Aalbserpin-1_2 protein was purified and obtained. In addition, we also found Aalbserpin-1_2 was expressed higher in salivary gland and midgut than in fat body (P<0.05). The results suggest that the Aalbserpin-1 gene from Ae. albopictus exists two alternative transcripts, and Aalbserpin-1_2 is expressed highly in salivary gland and midgut, indicating that the rAalbserpin-1_2 protein has been prokaryotic expressed and purified successfully.
程金芝, 孙宇, 陈璐, 刘鉴, 吴家红. 白纹伊蚊唾液腺serpin1基因的可变剪接分析与原核表达[J]. 中国人兽共患病学报, 2014, 30(5): 473-478.
CHENG Jin-zhi, SUN Yu, CHEN Lu, LIU Jian, WU Jia-hong. Alternative splicing and expression of the Alboserpin-1 from salivary gland of Aedes albopictus. Chinese Journal of Zoonoses, 2014, 30(5): 473-478.
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2014, Vol. 30 
