Zhang Yi-Ling, Xue Ren-Yu, Cao Guang-Li, Meng Xiang-Kun, Zhu Yue-Xiong, Pan Zhong-Hua, Gong Cheng-Liang
School of Biology & Basic Medical Sciences, Soochow University, No. 199 Ren'ai Road, Dushu Lake Higher Education Town, Suzhou Industrial Park, Suzhou, 215123, People's Republic of China,
Mol Biol Rep. 2014;41(4):2657-66. doi: 10.1007/s11033-014-3124-7. Epub 2014 Jan 28.
To explore whether the nonvirus encoded protein could be embedded into Bombyx mori cypovirus (BmCPV) polyhedra. The stable transformants of BmN cells expressing a polyhedrin (Polh) gene of BmCPV were constructed by transfection with a non-transposon derived vector containing a polh gene. The polyhedra were purified from the midguts of BmCPV-infected silkworms and the transformed BmN cells, respectively. The proteins embedded into polyhedra were determined by mass spectrometry analysis. Host derived proteins were detected in the purified polyhedra. Analysis of structure and hydrophilicity of embedded proteins indicated that the hydrophilic proteins, in structure, were similar to the left-handed structure of polyhedrin or the N-terminal domain of BmCPV structural protein VP3, which were easily embedded into the BmCPV polyhedra. The lysate of polyhedra purified from the infected transformation of BmN cells with modified B. mori baculovirus BmPAK6 could infect BmN cells, indicating that B. mori baculovirus could be embedded into BmCPV polyhedra. Both the purified polyhedra and its lysate could be coloured by X-gal, indicating that the β-galactosidase expressed by BmPAK6 could be incorporated into BmCPV polyhedra. These results suggested that some heterologous proteins and baculovirus could be embedded into polyhedra in an unknown manner.
为探究非病毒编码蛋白是否可嵌入家蚕质型多角体病毒(BmCPV)多角体中。通过用含有多角体蛋白(Polh)基因的非转座子衍生载体转染,构建了表达BmCPV多角体蛋白基因的BmN细胞稳定转化体。分别从感染BmCPV的家蚕中肠和转化的BmN细胞中纯化多角体。通过质谱分析确定嵌入多角体的蛋白。在纯化的多角体中检测到宿主来源的蛋白。对嵌入蛋白的结构和亲水性分析表明,亲水性蛋白在结构上类似于多角体蛋白的左手结构或BmCPV结构蛋白VP3的N端结构域,它们易于嵌入BmCPV多角体中。用修饰的家蚕杆状病毒BmPAK6感染转化的BmN细胞后纯化的多角体裂解物可感染BmN细胞,这表明家蚕杆状病毒可嵌入BmCPV多角体中。纯化的多角体及其裂解物均可被X-gal染色,这表明BmPAK6表达的β-半乳糖苷酶可掺入BmCPV多角体中。这些结果表明,一些异源蛋白和杆状病毒可以以未知方式嵌入多角体中。