Secretory Expression of Salivary ATP Diphosphohydrolase(Apyrase)from Aedes albopictus in Pichia pastoris

Abstract

Abstract: The gene-coding mature apyrase protein from Aedes albopictus was amplified by RT-PCR and cloned in frame with the α-factor secretion signal peptide into Pichia pastoris secreting expression vector pGAPZα-A resulting in the pGAPZα-A-apyrase. After being linearized by BlnⅠ restriction enzyme， the recombinant pGAPZα-A-apyrase was trans-formed into Pichia pastoris GS115 by electroporation. Recombinant strains pGAPZα-A-apyrase/GS115 were screened on YPDS plates containing Zeocin and identified by PCR. The recombinant protein of apyrase (Mr 60 000) has been expressed in the supernatant of Pichia pastoris.

Key words: Aedes albopictus, Apyrase, Pichia pastoris, Secreted expression

WU Song-quan1 *，WANG Guang-li1，LEI Yong-liang2，ZHANG Kai-bo1. Secretory Expression of Salivary ATP Diphosphohydrolase(Apyrase)from Aedes albopictus in Pichia pastoris[J]. , 2013, 31(1): 19-73-75.

[1]	WANG Guan-xi, LI Ya-shu, LI Yue-yue, CAO Yuan-yuan, YANG Meng-meng, ZHANG Mei-hua, WU Jing-yao, LIANG Cheng, LI Ju-lin, ZHOU Hua-yun, TANG Jian-xia, ZHU Guo-ding. Resistance to deltamethrin and knockdown resistance mutation in Aedes albopictus from Jiangsu Province [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022, 40(4): 468-474.
[2]	XIA Dan，TENG Ping-ying，CHEN Xiao-guang，ZHOU Xiao-hong*. Diapause of Aedes albopictus and the Related Molecular Mechanisms [J]. , 2016, 34(3): 18-282-289.
[3]	LI Xing-Pan, CAO Guo-Mei, XING Cui-Cui, HUA Qian-Qian, ZHANG Liang-Liang, LIANG Shao-Hui. Preparation and Antigenicity Analysis of Recombinant Aegyptin-like Protein of Aedes albopictus [J]. , 2014, 32(3): 6-193-197.
[4]	WU Song-quan1 *，WANG Guang-li1，ZHOU Wu1，ZHONG Shi-gen2. Cloning and Expression of Transferrin Protein from Culex pipiens pallens and a Study of its Antimicrobial Activity [J]. , 2014, 32(1): 8-38-41.
[5]	LIU Jian1，CHENG Jin-zhi1，SUN Yu2，ZHU Ru-fang3，ZHANG Liang4，WU Jia-hong1 *. Cloning and Expression of the Mucin-Related Protein1（Aamucin1）from Salivary Gland of Aedes albopictus [J]. , 2013, 31(2): 8-120-123.
[6]	CAI Yan-li，ZHENG Xue-li*. Cloning of AGO2 and Dcr-2 Gene Fragments and Analysis of Their Transcription Level in Different Developmental Stages of Aedes albopictus [J]. , 2012, 30(3): 10-214-217.
[7]	LIXiao-bo;WANGTing-ting;MAYan;ZHUJia-yong;LIUMan-yu;JINXiao-bao. Heterologous expression of attacin gene from Musca domestica* in Pichia pastoris [J]. , 2010, 28(5): 3-336.
[8]	WUJia-hong;CHENGJin-zhi;CHENLu;SUNYu. Expression of the Genes of Adenosine Deaminase, C-lectin andSerpin in the Salivary Gland of Aedes albopictus [J]. , 2010, 28(3): 7-193.
[9]	ZHANGLing-min;SUNJia-mei;LVHui-fang;SHIHai-ying. Effect of Alpha-terthienyl on Protein，Esterase andLipid Peroxidation of Aedes albopictus* Larvae [J]. , 2010, 28(1): 11-49.
[10]	GUJin-bao;SUNYan-tao;PENGHong-juan. Construction of a Yeast Model for Screening Aedes albopictus Ecdysone Agonist Pesticides [J]. , 2007, 25(5): 5-380.
[11]	LIUZhi-gang;HEYi-hua. Expression of Allergen Bla g 2 from Blattella germanica in Pichia pastoris [J]. , 2007, 25(2): 9-123.
[12]	ZHENGMei-juan;LIMin;WANGZhi-cheng;LUOFei;LUOQing-li;CHUDe-yong;LICong-lei;SHENJi-long. Secreted Expression of Signaling Protein 14-3-3 of Schistosoma japanicom in Pichia pastoris System with Primary Evaluation on its Antigenicity [J]. , 2007, 25(1): 3-16.
[13]	WUJia-hong;ZHAOTong-yan;DONGYan-de. Construction of Suppression Subtracted cDNA Library of Deltamethrin-resistant Aedes albopictus [J]. , 2006, 24(4): 10-284.
[14]	XIAOJing-ying;ZHANGDong-mei;CAILian-shun;SHENLu-hui;PANWei-qing. Evaluation of Immunogenicity and Protection Efficacy of the Recombinant Hypoxanthine-Guanine-Xanthine of Plasmodium falciparum in Mice [J]. , 2006, 24(3): 8-195.
[15]	WANGShao-hua;ZHUXin-ping;GUYuan;YANGYa-ping;YANGJing. Construction and Expression of the Ts87 Gene of Trichinella spiralis* in Pichia pastoris [J]. , 2005, 23(4): 12-239.

Secretory Expression of Salivary ATP Diphosphohydrolase(Apyrase)from Aedes albopictus in Pichia pastoris

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments