家蚕血细胞特异表达新基因Bm04862地鉴定及表达.PDFVIP

生 物 工 程 学 报 余霜 等/家蚕血细胞特异表达新基因Bm04862 的鉴定及表达 241 Chinese Journal of Biotechnology /cjbcn February 25, 2016, 32(2): 241−249 DOI: 10.13345/j.cjb.150115 ©2016 Chin J Biotech, All rights reserved 生物技术与方法 Bm04862 余霜，苏晶晶，徐曼，张奎，崔红娟 400716 , , , . Bm04862 . , 2016, 32(2): 241–249. Yu S, Su JJ, Xu M, et al. Identification and characterization of a novel hemocyte specific gene Bm04862 in silkworm, Bombyx mori. Chin J Biotech, 2016, 32(2): 241–249. 摘 要: 通过家蚕组织芯片数据筛选得到家蚕血细胞特异表达基因 Bm04862 ，并首次对该基因进行了克隆与 鉴定。应用RACE 技术获得该基因全长，并对其进行生物信息学分析。Bm4862 基因开放阅读框 819 bp ，共编 码273 个氨基酸残基，预测其为跨膜蛋白；通过qRT-PCR 技术对其时空表达情况进行分析；结果显示Bm04862 基因在家蚕血细胞中特异高表达，并在 4 龄眠期和预蛹 2 d 时达到表达高峰；构建Bm04862 真核表达载体， 转染Sf9 细胞分析其蛋白的亚细胞定位情况，结果表明其定位于细胞核膜和部分细胞质中。此外，用大肠杆菌 刺激蚕体24 h 后，Bm04862 基因表达水平显著上调，表明大肠杆菌可以诱导该基因的表达，由此推测该基因 可能参与家蚕的免疫应答。这为深入研究该基因在家蚕免疫反应中的功能提供了参考。 : 家蚕，血细胞特异基因，Bm04862 ，表达特征，亚细胞定位 Identification and characterization of a novel hemocyte specific gene Bm04862 in silkworm, Bombyx mori Shuang Yu, Jingjing Su, Man Xu, Kui Zhang, and Hongjuan Cui Southwest University, State Key Laboratory of Silkworm Genome Biology, Chongqing 400716, China Abstract: Bm04862 is a novel hemocyte-specific gene, which has been identified and cloned through the microarray data in silkworm, Bombyx mori . Initially, we successfully obtained the full-length cDNA sequence of Bm04862 via the rapid amplification of cDNA ends (RACE). The sequence consists of an open reading frame (ORF) with 819 bp, which could encode 273 amino acid residues. The bioinformatics analysis predicted that Bm04862 was a transmembrane protein. Received: March