| 鲢IL-8基因的克隆与表达分析 |
| |
| 引用本文： | 孙军,梁卉,林小涛,胡云凤,童芳.鲢IL-8基因的克隆与表达分析[J].淡水渔业,2009,39(2). |
| |
| 作者姓名： | 孙军 梁卉 林小涛 胡云凤 童芳 |
| |
| 作者单位： | 暨南大学水生生物研究所,广州,510632 |
| |
| 基金项目： | 国家自然科学基金，广东省联合基金重点项目 |
| |
| 摘 要： | 采用RACE-PCR方法,从鲢(Hypophthalmichthys molitrix)头肾SMARTcDNA中获得了679 bp IL-8 cDNA序列。结果显示:该序列包含169 bp的5′非编码区,213 bp 3′端非编码区和297 bp的开放阅读框;鲢IL-8的开放阅读框编码98个氨基酸残基,其中包含构成两对二硫键的4个保守半胱氨酸。RT-PCR结果显示鲢:IL-8 mRNA主要在胰、肝脏和头肾中表达。将鲢IL-8不含信号肽的成熟多肽克隆到原核融合表达载体pQE30上,转入大肠杆菌M15内进行表达,经SDS-PAGE电泳后显示,重组融合蛋白在分子量约11kD处有一明显表达带,与预期计算的分子量大小相一致。
|
| 关 键 词： | 鲢(Hypophthalmichthys molitrix) IL-8 克隆 组织表达 原核表达 |
Gene cloning and expression of Interleukin-8(IL-8) from silver carp(Hypophthalmichthys molitrix) |
| |
| SUN Jun,LIANG Hui,LIN Xiao-tao,HU Yun-feng,TONG Fang.Gene cloning and expression of Interleukin-8(IL-8) from silver carp(Hypophthalmichthys molitrix)[J].Freshwater Fisheries,2009,39(2). |
| |
| Authors: | SUN Jun LIANG Hui LIN Xiao-tao HU Yun-feng TONG Fang |
| |
| Institution: | SUN Jun,LIANG Hui,LIN Xiao-tao,HU Yun-feng,TONG Fang(Institute of Hydrobiology,Jinan University,Guangzhou 510632) |
| |
| Abstract: | Interleukin-8(IL-8) is a CXC chemokine,which can attract and activate specific types of leukocytes,such as lymphocytes and neut-rophils.The cDNA of silver carp IL-8 consists of 679 bp with 169 bp 5′ untranslated region(UTR),213 bp 3′UTR.An open reading frame of 297 bp encoded a 98-amino acid peptide,with four cysteines.Phylogenetic analyses clearly demonstrated that silver carp IL-8 resembles the IL-8s of other fish.RT-PCR results revealed that stronger products in the pancreas,liver and head kidney were detected.Then the mature peptide of silver carp IL-8 was directionally connected the expression vector pQE30 to construct pQE30-IL8 recombinant.This recombinant was transformed into E.coli M15.An expected protein about 11kD was found in M15 cell protein after inducting with IPTG by SDS-PAGE analysis. |
| |
| Keywords: | IL-8 |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
