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家蚕感染质型多角体病毒相关基因Arylphor in的克隆与鉴定
  • ISSN号:0578-1752
  • 期刊名称:《中国农业科学》
  • 时间:0
  • 分类:S763.421[农业科学—森林保护学;农业科学—林学]
  • 作者机构:[1]江苏科技大学生物与化学工程学院,江苏镇江212018, [2]江苏科技大学蚕业研究所,江苏镇江212018, [3]中国农业科学院蚕业研究所,江苏镇江212018
  • 相关基金:基金项目:国家自然科学基金(30972143)、江苏科技大学博士启动基金(635211206)
中文摘要:

【目的】克隆家蚕Arylphorin的cDNA全长序列,利用生物信息学对该序列进行分析,检测该基因在家蚕感染质型多角体病毒(Bombyx mori cytopiasmi cpolyhedrosis virus,BmCPV)后的表达量变化模式,为进一步研究其生物学功能奠定基础。【方法】以家蚕中肠总RNA为模板反转录合成cDNA,利用cDNA末端快速扩增(RACE)技术克隆家蚕Arylphorin的全长cDNA;通过实时荧光定量PCR(qRT-PCR)分析其组织表达模式和在家蚕感染BmCPV后的表达差异。【结果】克隆得到了家蚕Arylphorin的全长cDNA(命名为BmAryl),在GenBank登录号为JN581664。该基因的cDNA全长为2260bp,包含1个26bp5啡翻译区和1个143bp的3啡翻译区,3’非翻译区包含有终止密码子、多聚腺嘌呤信号AATAAA和poly(A)尾。其最大开放阅读框(ORF)为2091bp,编码的蛋白由696个氨基酸组成,预测蛋白分子量为82.8kD,等电点为6.07。多序列比对和系统进化分析发现,家蚕BmAryl蛋白与家蚕的性别专一贮藏蛋白2相似性最高,为66%。Bmhryl主要在家蚕的脂肪体和血淋巴中表达,精巢和卵巢中的表达量无明显差别;而在家蚕感染BmCPV后该基因在中肠的表达量明显下调。【结论】成功克隆获得BmAryl的全长cDNA,其编码的蛋白质属于芳香蛋白家族,BmAryl主要在家蚕的脂肪体和血淋巴中表达,不存在性别专一性,且家蚕感染BmCPV后BmAryl在中肠的表达明显下调。

英文摘要:

[Objective] The objective of this study is to clone the full-length cDNA sequence of Arylphorin in silkworm (Bombyx mori) to analyze the gene sequence by bioinformatics and to investigate the mRNA expression level after the B. mori infected by cytoplasmic polyhedrosis virus (BmCPV), and to provide a new foundation for the further study of its biological function. [ Method ] Rapid amplification of cDNA ends (RACE) approach was employed to clone the full-length cDNA of Arylphorin from the total RNA of the silkworm midgut. Fluorescent quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze the expression profiling of Arylphorin in different tissues and at different time points after the BmCPV infection. [ Result ] The full-length cDNA of Arylphorin was obtained and named as BmAryl with accession number of JN581664 in GenBank. It consisted of a 5'-terminal untranslated region (UTR) of 26 bp and a 3'-UTR of 143 bp with polyadenylation signal sequences AATAAA and a poly (A) tail. The longest open reading frame (ORF) encoded a polypeptide of 696 amino acids with a theoretical isoelectric point of 6.07and the predicted molecular weight of 82.8 kD. Sequence comparison showed that BmAryl was 66% identical to B. mori sex-specific storage-protein 2 precursor, mRNA transcripts of BmAryl were detected mainly in fat body and hemolymph and the expression level had no difference between the testicle and ovary. For the silkworm larvae infected by BmCPV, the relative expression level of BmAryl was significantly down-regulated in the midgut. [ Conclusion] The full-length cDNA of BmAryl was successfully cloned from silkworm, which encodes a protein that belongs to the aromatic protein family. BmAryl expression were detected mainly in fat body and hemolymph, not sex-specific but down-regulated in the midgut due to BmCPV infection. These results has provided not only helpful information for further studying the function of BmAryl in silkworm but also be helpful for understanding o

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期刊信息
  • 《中国农业科学》
  • 中国科技核心期刊
  • 主管单位:中华人民共和国农业部
  • 主办单位:中国农业科学院 中国农学会
  • 主编:万建民
  • 地址:北京中关村南大街12号中国农业科学院图书馆楼4101-4103室
  • 邮编:100081
  • 邮箱:zgnykx@caas.cn
  • 电话:010-82109808 82106279
  • 国际标准刊号:ISSN:0578-1752
  • 国内统一刊号:ISSN:11-1328/S
  • 邮发代号:2-138
  • 获奖情况:
  • 中国期刊方阵“双高”期刊,第三届中国出版政府奖提名奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),英国食品科技文摘,中国北大核心期刊(2000版)
  • 被引量:85620