中国海洋大学学报（自然科学版）
中國海洋大學學報（自然科學版）
중국해양대학학보（자연과학판）
PERIODICAL OF OCEAN UNIVERSITY OF CHINA
2010年
4期
31-37
,共7页
王贤丽%张玉喜%孟亮%陈松林
王賢麗%張玉喜%孟亮%陳鬆林
왕현려%장옥희%맹량%진송림
大菱鲆%IgL%cDNA%感染%RT-PCR
大蔆鲆%IgL%cDNA%感染%RT-PCR
대릉평%IgL%cDNA%감염%RT-PCR
turbot(Scophthalmus maximus)%IgL%cDNA%infection%RT-PCR
实验采用末端快速扩增(RACE)技术从大菱鲆脾脏cDNA文库中筛选得到了免疫球蛋白轻链IgL的全长cDNA片段.该序列包含47 bp的5'末端非编码区(5'UTR),738 bp的开放阅读框(ORF)和202 bp 3'UTR,整个开放阅读框编码246个氨基酸.系统发生分析表明大菱鲆IgL基因与五条鰤的IgL基因起源关系最近.RT-PCR分析表明,大菱鲆IgL基因只在正常脾脏、肾脏和头肾组织中表达;IgL在大菱鲆胚胎发育细胞期就已开始表达,在大菱鲆胚胎尾芽期和体节期表达持续增强;在鳗弧菌感染大菱鲆脾脏和头肾早期均检测到IgL基因强烈表达,后期表达逐渐减弱;大菱鲆胚胎细胞(TEC)在用鳗弧菌感染48h后,IgL开始表达.这些结果均表明IgL基因在大菱鲆免疫应答中起着重要作用.
實驗採用末耑快速擴增(RACE)技術從大蔆鲆脾髒cDNA文庫中篩選得到瞭免疫毬蛋白輕鏈IgL的全長cDNA片段.該序列包含47 bp的5'末耑非編碼區(5'UTR),738 bp的開放閱讀框(ORF)和202 bp 3'UTR,整箇開放閱讀框編碼246箇氨基痠.繫統髮生分析錶明大蔆鲆IgL基因與五條鰤的IgL基因起源關繫最近.RT-PCR分析錶明,大蔆鲆IgL基因隻在正常脾髒、腎髒和頭腎組織中錶達;IgL在大蔆鲆胚胎髮育細胞期就已開始錶達,在大蔆鲆胚胎尾芽期和體節期錶達持續增彊;在鰻弧菌感染大蔆鲆脾髒和頭腎早期均檢測到IgL基因彊烈錶達,後期錶達逐漸減弱;大蔆鲆胚胎細胞(TEC)在用鰻弧菌感染48h後,IgL開始錶達.這些結果均錶明IgL基因在大蔆鲆免疫應答中起著重要作用.
실험채용말단쾌속확증(RACE)기술종대릉평비장cDNA문고중사선득도료면역구단백경련IgL적전장cDNA편단.해서렬포함47 bp적5'말단비편마구(5'UTR),738 bp적개방열독광(ORF)화202 bp 3'UTR,정개개방열독광편마246개안기산.계통발생분석표명대릉평IgL기인여오조사적IgL기인기원관계최근.RT-PCR분석표명,대릉평IgL기인지재정상비장、신장화두신조직중표체;IgL재대릉평배태발육세포기취이개시표체,재대릉평배태미아기화체절기표체지속증강;재만호균감염대릉평비장화두신조기균검측도IgL기인강렬표체,후기표체축점감약;대릉평배태세포(TEC)재용만호균감염48h후,IgL개시표체.저사결과균표명IgL기인재대릉평면역응답중기착중요작용.
A novel turbot (Scophthalmus maximus) immunoglobulin light chain (IgL) was screened from a turbot spleen cDNA library. The complete cDNA of the turbot IgL contains a 47 bp 5' UTR, a 738 bp open reading frame (ORF) encoding 246 amino acids and a 202 bp 3'UTR. Phylogenetic analysis showed that the turbot IgL clustered with S.quinqueradiata IgL. RT-PCR demonstrated that turbot IgL was expressed in spleen, kidney and head kidney from uninfected adults. IgL expressed during the early stages of embryo development and gradually increased until larva stage. The expression of IgL was also dramatically increased after challenge in turbot spleen and head kidney after 5 h. Furthermore, the turbot LCK was induced after 48 h in turbot embryonic cells (TECs) after challenge with Vibrio anguillarum. These results indicated that the turbot LCK played an important role in turbot immune response.
