| 摘要: |
| 从前期构建的短蛸(Octopus ocellatus)cDNA 文库中克隆获得了一个短蛸酪氨酸蛋白激酶(OoBtk)基因的cDNA 全长, 其cDNA 全长1191 bp, 包括5'非编码区(UTR)259 bp, 3'UTR 227 bp, 开放阅读框(ORF)705 bp, 编码234 个氨基酸, 预测理论等电点为8.50, 分子量为27.7 kDa。运用实时荧光定量PCR法分析了在健康短蛸的不同组织以及鳗弧菌和藤黄微球菌刺激后血细胞中OoBtk 的表达规律, 结果表明: OoBtk 在肌肉、外套膜、鳃、鳃心、系统心脏、肝胰脏、肾囊、胃、性腺和血细胞等各检测组织中均有表达, 其中在肝胰脏的表达量最高; 短蛸经鳗弧菌和藤黄微球菌刺激后,OoBtk 在血细胞中的表达量呈现出了明显的被诱导表达趋势, 在鳗弧菌和藤黄微球菌刺激后6 h 表达即增强, 并分别在鳗弧菌刺激后48 h 和藤黄微球菌刺激后12 h 达到最高。OoBtk 参与短蛸对于鳗弧菌和藤黄微球菌刺激的应答过程。 |
| 关键词: 短蛸 酪氨酸蛋白激酶 实时荧光定量PCR 藤黄微球菌 鳗弧菌 |
| DOI：10.11693/hyhz20140900240 |
| 分类号: |
| 基金项目:国家自然科学基金项目, 31272643号, 31302215号; 山东省自然科学基金项目, ZR2013CQ030号; 烟台市科技发展计划项目, 2011068号; 水生动物营养与饲料“泰山学者”岗位资助。 |
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| CLONING OF A TYROSINE PROTEIN KINASE GENE FROM OCTOPUS OCELLATUS AND THE EXPRESSION PATTERN AFTER BACTERIA STIMULATION |
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ZHANG Ran-Ran1,2, WEI Xiu-Mei2, YANG Jian-Min2, WANG Wei-Jun2, LIU Xiang-Quan2, FENG Yan-Wei2
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1.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;2.Shandong Provincial Key Laboratory of Restoration for Marine Ecology, Shandong Marine Resource and Environment Research Institute, Yantai 264006, China
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| Abstract: |
| A full-length cDNA encoding OoBtk was cloned from the cDNA Library of Octopus ocellatus in this study. It was 1191 bp, containing a 5' untranslated region of 259 bp, a 3'UTR of 277 bp, and an open reading frame (ORF) of 705 bp encoding a polypeptide of 234 amino acids with an isoelectric point of 8.50 and predicted molecular weight of 27.7 kDa. The mRNA expression patterns of an OoBtk in tissues and haemocytes after Listonella anguillarum or Micrococcus luteus challenge were characterized by real-time PCR. The results show that OoBtk was widely expressed in muscle, mantle, gill, branchial heart, systemic heart, hepatopancreas, saccus renalis, stomach, gonad, and hemocyte, highest in hepatopancreas. The mRNA expression of OoBtk in hemocytes showed an obviously induced trend after L. anguillarum or M. luteus stimulation; it was significantly up-regulated at 6h post stimulation, and peaked at 48h or 12h post stimulation, respectively. Therefore, OoBtk in O. ocellatus is involved in the response against L. anguillarum or M. luteus challenge. |
| Key words: Octopus ocellatus tyrosine protein kinase real-time PCR Listonella anguillarum Micrococcus luteus |
