cDNA cloning and characterization of LASP1 from silkworm, Bombyx mori, involved in cytoplasmic polyhedrosis virus infection
cDNA cloning and characterization of LASP1 from silkworm, Bombyx mori, involved in cytoplasmic polyhedrosis virus infection
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DOI：
10.1016/j.gene.2012.09.062
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发表时间：
2012-12-15
期刊：
影响因子：
3.5
通讯作者：
Guo, Xi-jie
中科院分区：
文献类型：
--
作者：
Gao, Kun;Deng, Xiang-yuan;Guo, Xi-jie
通过快速扩增 cDNA 末端，首次从家蚕 Bombyx mori 中鉴定出含有 LIM 和 SH3 的蛋白 1（命名为 BmLASP1）的全长 cDNA。 BmLASP1 cDNA 全长 2094 bp，由 117 bp 的 5' 端非翻译区 (UTR) 和 610 bp 的 3'-UTR 组成，其中包含两个聚腺苷酸化信号序列 AATAAA 和一个聚 (A) 尾。 BmLASP1 cDNA编码包含455个氨基酸的多肽，包括LIM结构域、两个nebulin结构域和SH3结构域。理论等电点为7.07，预测分子量为51.8 kDa。 BmLASP1 没有信号肽，但有三个潜在的 N-糖基化位点。序列相似性和系统发育分析表明，BmLASP1属于昆虫LASP1类群，其连接子区域较长，与脊椎动物LASP1不同。家蚕的LASP1编码区含有8个外显子，最后一个外显子-内含子边界与哺乳动物和玻璃海鞘LASP1基因一样保守。荧光定量实时聚合酶链式反应显示BmLASP1 mRNA转录本主要在性腺、头部、气门中检测到，丝腺、血管粘膜、中肠、脂肪体、血细胞中也有少量表达。家蚕幼虫感染家蚕胞质多角体病毒（BmCPV）后，中肠中BmLASP1的相对表达水平下调。这一结果表明BmLASP1可能在家蚕对BmCPV感染的反应中发挥重要作用。 (C) 2012 Elsevier B.V. 保留所有权利。
Full-length cDNA of a LIM and SH3 contained protein 1 (named BmLASP1) was identified from the silkworm, Bombyx mori, for the first time by rapid amplification of cDNA ends. The full-length cDNA of BmLASP1 is 2094 bp, consisting of a 5'-terminal untranslated region (UTR) of 117 bp, and a 3'-UTR of 610 bp with two poly-adenylation signal sequence AATAAA and a poly (A) tail. The BmLASP1 cDNA encodes a polypeptide comprising 455 amino acids, including a LIM domain, two nebulin domains and an SH3 domain. The theoretical isoelectric point is 7.07 and the predicted molecular weight is 51.8 kDa. BmLASP1 has no signal peptide but three potential N-glycosylation sites. Sequence similarity and phylogenic analyses indicated that BmLASP1 belonged to the group of insect LASP1 with a longer linker region which is different from vertebrate LASP1. The LASP1 in silkworm contained eight exons in its coding regions, and the last exon-intron boundary was conserved the same as in mammalian and Ciona intestinalis LASP1 genes. By fluorescent quantitative real-time polymerase chain reaction, the mRNA transcripts of BmLASP1 were mainly detected in the gonad, head, and spiracle, and slightly in the silk gland, vasa mucosa, midgut, fat body, and hemocytes. After silkworm larvae were infected by B. mori cytoplasmic polyhedrosis virus (BmCPV), the relative expression level of BmLASP1 was down-regulated in the midgut. This result suggested that BmLASP1 may play an important role in the response of silkworm to BmCPV infection. (C) 2012 Elsevier B.V. All rights reserved.