Construction of insect expression vector pIB/V5-His of AcerOr2 in Apis cerana cerana

doi:10.3969/j.issn.1004-1524.2020.06.07

›› 2020, Vol. 32 ›› Issue (6): 994-999.DOI: 10.3969/j.issn.1004-1524.2020.06.07

• Animal Science • Previous Articles Next Articles

Construction of insect expression vector pIB/V5-His of AcerOr2 in Apis cerana cerana

GUO Lina¹, ZHAO Huiting², REN Youshe¹, XU Bing³, JIANG Yusuo^1,*

1. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China;
2. College of Life Science, Shanxi Agricultural University, Taigu 030801, China;
3. College of Engineering, Shanxi Agricultural University, Taigu 030801, China

Received:2019-12-05 Online:2020-06-25 Published:2020-06-24

Abstract

Abstract: In order to construct a recombinant expression vector which can stably and efficiently express olfactory receptor AcerOr2 in Apis cerana cerana, and then express in the Sf9 insect cell, AcerOr2 was amplified and the insect expression vector pIB/V5-His were treated with BamHⅠ and EcoRⅠ, and AcerOr2 were ligated into pIB/V5-His vector using T4 DNA ligasein this experiment. The recombinant DNA was transfected into Sf9 cells by liposome transfection, and the expression and subcellular localization of AcerOr2 protein were detected by Western blot and immunofluorescence. The results showed that the recombinant expression vector pIB/V5-His-AcerOr2 was successfully constructed and a stable transfected vector was expressed. Western blot results showed that the recombinant expression vector could be expressed in the insect cell Sf9, the fusion protein was about 56 ku. Immunofluorescence showed that the recombinant expression vector was localized in the Sf9 cell membrane, which was consistent with the predicted results. The results showed that the construction of recombinant expression vector pIB/V5-His-AcerOr2 could stably express in Sf9 cell, which will lay a foundation for the further study on the function of AcerOr2 in Apis cerana cerana.

Key words: Apis cerana cerana, olfactory receptor AcerOr2, vector construction, subcellular localization

CLC Number:

S891

GUO Lina, ZHAO Huiting, REN Youshe, XU Bing, JIANG Yusuo. Construction of insect expression vector pIB/V5-His of AcerOr2 in Apis cerana cerana[J]. , 2020, 32(6): 994-999.

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Construction of insect expression vector pIB/V5-His of AcerOr2 in Apis cerana cerana

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