Expression of the melittin gene of Apis cerana cerana (Hymenoptera: Apidae) in insect cells

Title:

Expression of the melittin gene of Apis cerana cerana (Hymenoptera: Apidae) in insect cells

Creator:

Shi, Wan-Jun, Cheng, Jia-An, and Zhang, Chuan-Xi

Identifier:

https://cdk.lib.cas.cz/client/handle/uuid:ac78e425-3f5d-4b80-a5bc-2b859a257cae
uuid:ac78e425-3f5d-4b80-a5bc-2b859a257cae

Subject:

Hymenoptera, Apidae, Apis cerana cerana, Chinese honeybee, melittin, insect cells, and protein expression

Type:

article, model:article, and TEXT

Description:

A fragment encoding melittin cDNA from Apis cerana cerana fused with glutathione S-transferase gene was inserted into the multiple cloning site of the pBacFastHTb to construct a recombinant donor plasmid, pBacHT-GSTAccM, which was transposed to the target bacmid in E. coli (DH10) by Tn7 transposition function. Then the recombinant baculovirus Bacmid-GSTAccM was transfected into Tn-5B1-4 cells of the cabbage looper, Trichoplusia ni, mediated by lipofectin. The expressed protein of about 34 kDa was detected by Western blotting and triple antibody sandwich ELISA, indicating that the recombinant protein is the fusion protein of GSTAccM. Thin layer scanning showed that the expression level of GSTAccM was about 7% of the total cell protein. Purified and recovered recombinant melittin of A. c. cerana showed bioactivity in activating rabbit platelets to aggregate.

Language:

English

Rights:

http://creativecommons.org/publicdomain/mark/1.0/
policy:public

Source:

European Journal of Entomology | 2006 Volume:103 | Number:4

Harvested from:

CDK

Metadata only:

false