Expression and biological function identification of TCRγ9/δ2-Fc protein in baculovirus vector expression system
- VernacularTitle:昆虫杆状病毒表达系统表达TCRγ9/δ2-Fc融合蛋白及其生物学功能的初步鉴定
- Author:
Yang GUO
;
Jing ZHENG
;
Yu HU
;
Lianxian CUI
;
Wei HE
- Publication Type:Journal Article
- Keywords:
TCRγ9/δ2-Fc;
baculovirus vector expression system
- From:
Basic & Clinical Medicine
2009;29(12):1268-1272
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an expression system of TCRγ9/δ2-Fc protein by baculovirus vector ex-pression system and identify biological function of expressed TCRγ9/δ2-Fc protein. Methods γ9Fc and 82 (OT3) Fc gene fragments were amplified by overlap PCR and inserted into expression vector pBACp10ph. The recombinant plasmid pBACp10ph-γ9/δ2(OT3)-Fc and the baculovirus DNA were co-transfected into st9 cells. The expression position of TCRγ9/δ2 (OT3)-Fc was identified by Western blot and the expression efficiency of γ9Fc and δ2 (OT3) Fc was tested by flow cytometry (FCM). Furthermore, the binding activity of TCRγ9/δ2 (OT3)-Fc protein with SKOV3 ceils and MNS protein was evaluated with laser scanning confocal microslopy and surface plasmon resonance (SPR). Results The recombinant vector pBACp10ph-γ9/δ2(OT3)-Fc was constructed and TCRγ9/δ2(OT3)-Fc protein was expressed in sf9 ceils. However, the expression efficiency of γ9Fc and 82 (0T3) Fc was quite differ-ent. It was proved that purified TCRγ9/δ2 (OT3)-Fc protein can bind with SKOV3 cell and MNS protein. Conclu-sion TCRγ9/δ2-Fc protein is successfully expressed in baculovirus vector expression system and TCRγ9/δ2-Fc protein can simulate the binding activity of TCR in vitro.
