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家蚕Bmserpin-2基因的分子克隆、表达及特性分析

Molecular cloning, expression and characterization of Bmserpin-2 gene from Bombyx mori.

作者信息

Pan Ye, Xia Hengchuan, Lü Peng, Chen KePing, Yao Qin, Chen Huiqin, Gao Lu, He Yuanqing, Wang Lin

机构信息

Institute of Life Sciences, Jiangsu University, Zhenjiang, PR China.

出版信息

Acta Biochim Pol. 2009;56(4):671-7. Epub 2009 Dec 7.

Abstract

Serpins are a broadly distributed family of protease inhibitors. In this study, the gene encoding Bombyx mori serpin-2 (Bmserpin-2) was cloned and expressed in E. coli. The Bmserpin-2 cDNA contains a 1125 bp open reading frame (ORF). The deduced protein has 374 amino-acid residues, contains a conserved SERPIN domain and shares extensive homology with other invertebrate serpins. RT-PCR analysis showed that Bmserpin-2 was expressed in all developmental stages of B. mori larvae and various larval tissues. Subcellular localization analysis indicated that Bmserpin-2 protein was located in the cytoplasm. Interestingly, real-time quantitative PCR revealed that the expression of Bmserpin-2 in the midgut of susceptible B. mori strain 306 significantly increased at 72 hours post inoculation (hpi) when infected with BmNPV. However, there was no significant increase of the Bmserpin-2 expression in resistant strain NB infected with BmNPV. Thus, our data indicates that Bmserpin-2 may be involved in B. mori antiviral response.

摘要

丝氨酸蛋白酶抑制剂(Serpins)是一类广泛分布的蛋白酶抑制剂家族。在本研究中,编码家蚕丝氨酸蛋白酶抑制剂-2(Bmserpin-2)的基因被克隆并在大肠杆菌中表达。Bmserpin-2 cDNA包含一个1125 bp的开放阅读框(ORF)。推导的蛋白质有374个氨基酸残基,包含一个保守的丝氨酸蛋白酶抑制剂结构域,并且与其他无脊椎动物的丝氨酸蛋白酶抑制剂有广泛的同源性。RT-PCR分析表明,Bmserpin-2在家蚕幼虫的所有发育阶段和各种幼虫组织中均有表达。亚细胞定位分析表明,Bmserpin-2蛋白位于细胞质中。有趣的是,实时定量PCR显示,在感染BmNPV后,易感家蚕品系306中肠内Bmserpin-2的表达在接种后72小时(hpi)显著增加。然而,在感染BmNPV的抗性品系NB中,Bmserpin-2的表达没有显著增加。因此,我们的数据表明,Bmserpin-2可能参与家蚕的抗病毒反应。

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