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海湾扇贝(Argopecten irradians,拉马克,1819年)中肽聚糖识别蛋白(PGRP)基因的分子克隆与mRNA表达

Molecular cloning and mRNA expression of peptidoglycan recognition protein (PGRP) gene in bay scallop (Argopecten irradians, Lamarck 1819).

作者信息

Ni Duojiao, Song Linsheng, Wu Longtao, Chang Yaqing, Yu Yundong, Qiu Limei, Wang Lingling

机构信息

Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.

出版信息

Dev Comp Immunol. 2007;31(6):548-58. doi: 10.1016/j.dci.2006.09.001. Epub 2006 Oct 4.

Abstract

Peptidoglycan recognition proteins (PGRPs) are a type of pattern recognition molecules (PRM) that recognize the unique cell wall component peptidoglycan (PGN) of bacteria and are involved in innate immunity. The first bivalve PGRP cDNA sequence was cloned from bay scallop Argopecten irradians by expressed sequence tag (EST) and PCR technique. The full-length cDNA of bay scallop PGRP (designated AiPGRP) gene contained 1018bp with a 615-bp open reading frame that encoded a polypeptide of 205 amino acids. The predicted amino acid sequence of AiPGRP shared high identity with PGRP in other organisms, such as PGRP precursor in Trichoplusia ni and PGRP SC2 in Drosophila melanogaster. A quantitative reverse transcriptase Real-Time PCR (qRT-PCR) assay was developed to assess the mRNA expression of AiPGRP in different tissues and the temporal expression of AiPGRP in the mixed primary cultured hemocytes challenged by microbial components lipopolyssacharide (LPS) from Escherichia coli and PGN from Micrococcus luteus. Higher-level mRNA expression of AiPGRP was detected in the tissues of hemocytes, gonad and kidney. The expression of AiPGRP in the mixed primary cultured hemocytes was up regulated after stimulated by PGN, while LPS from E. coli did not induce AiPGRP expression. The results indicated that AiPGRP was a constitutive and inducible expressed protein that was mainly induced by PGN and could be involved in scallop immune response against Gram-positive bacteria infection.

摘要

肽聚糖识别蛋白(PGRPs)是一类模式识别分子(PRM),可识别细菌独特的细胞壁成分肽聚糖(PGN),并参与固有免疫。通过表达序列标签(EST)和PCR技术,首次从海湾扇贝栉孔扇贝中克隆出双壳贝类PGRP cDNA序列。海湾扇贝PGRP(命名为AiPGRP)基因的全长cDNA包含1018bp,其中有一个615bp的开放阅读框,编码一个205个氨基酸的多肽。预测的AiPGRP氨基酸序列与其他生物中的PGRP具有高度同源性,如粉纹夜蛾中的PGRP前体和黑腹果蝇中的PGRP SC2。开发了一种定量逆转录实时PCR(qRT-PCR)分析方法,以评估AiPGRP在不同组织中的mRNA表达以及在受到来自大肠杆菌的微生物成分脂多糖(LPS)和来自藤黄微球菌的PGN刺激的混合原代培养血细胞中AiPGRP的时间表达。在血细胞、性腺和肾脏组织中检测到较高水平的AiPGRP mRNA表达。PGN刺激后,混合原代培养血细胞中AiPGRP的表达上调,而大肠杆菌的LPS未诱导AiPGRP表达。结果表明,AiPGRP是一种组成型且可诱导表达的蛋白,主要由PGN诱导,可能参与扇贝对革兰氏阳性菌感染的免疫反应。

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