Cao Qing, Zhu Shan-Ying, Wu Yan, Liu Yong, Zhu Jiang, Wang Wenbing
Institute of Life Science, Jiangsu University, Zhenjiang 212013, China.
School of The Environment, Jiangsu University, Zhenjiang 212013, China.
Pol J Microbiol. 2012 Sep 28;61(3):183-189.
The conotoxin-like (ctx) gene encodes a small cysteine-rich polypeptide in various baculoviruses. Previous research has demonstrated that the product of the ctx gene could be purified from insect cells infected by Autographa californica nuclear polyhedrosis virus (AcMNPV), but its function was unknown. In this paper, we compared the conserved cysteine motif structure (CX3GX2CX5CCX3CX6C) of the ctx gene in baculoviruses and generated recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV) with the BmNPV bacmid system. The recombinant BmNPV contained the ctx gene from AcMNPV or a fusion gene of ctx with eGFP, respectively. Fluorescence in CTX-eGFP-positive cells was mainly observed on the cell membrane. To gain insight into CTX function, two methods were used to elucidate the affect CTX had on hemolymph melanization in vivo and in vitro in insect larvae and pupae. The results indicated that CTX abrogates hemolymph melanization; however, the mechanisms require further evaluation.
芋螺毒素样(ctx)基因在多种杆状病毒中编码一种富含半胱氨酸的小多肽。先前的研究表明,ctx基因的产物可以从被苜蓿银纹夜蛾核型多角体病毒(AcMNPV)感染的昆虫细胞中纯化出来,但其功能尚不清楚。在本文中,我们比较了杆状病毒中ctx基因保守的半胱氨酸基序结构(CX3GX2CX5CCX3CX6C),并利用BmNPV杆粒系统构建了重组家蚕核型多角体病毒(BmNPV)。重组BmNPV分别包含来自AcMNPV的ctx基因或ctx与eGFP的融合基因。在CTX-eGFP阳性细胞中,荧光主要出现在细胞膜上。为了深入了解CTX的功能,我们采用了两种方法来阐明CTX对昆虫幼虫和蛹体内外血淋巴黑化的影响。结果表明,CTX可消除血淋巴黑化;然而,其机制需要进一步评估。