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Alternative TitleIdentification of a membrane protein gene from Rana Grylio Virus
Thesis Advisor张奇亚
Degree Grantor中国科学院水生生物研究所
Place of Conferral水生生物研究所
Keyword蛙虹彩病毒 Pcr 原核表达 Rt-pcr 亚细胞定位 Western Blot
Abstract从本实验室保存的蛙虹彩病毒(Ranivirus)RGV(Rana grylio virus)的基因组中,克隆出一个预测可能是膜蛋白的基因MMP (Myristylated membrane protein),运用病毒学技术、免疫学技术和分子生物学技术,对MMP基因进行特征分析。 根据与RGV同源性很高的FV3相应基因设计引物从RGV 基因组中克隆出全长1038bp 的MMP基因,对其测序并使用几种软件进行分析。MMP基因最大的ORF长972bp,编码323个氨基酸,分子量为35kDa,氨基酸序列与同属的其它病毒相应蛋白的同源性高达91-99%,预测氨基酸序列有一个跨膜区。 对MMP基因进行原核表达,得到分子量为53 kDa的缺失跨膜区的融合蛋白表达产物,制备出抗体。通过亚细胞定位和Western blot等实验初步证实MMP基因编码膜蛋白。
Other AbstractIn this study, the primers was designed according to MMP (Myristylated membrane protein)gene sequence of FV3, and then cloned the MMP gene using genome of RGV as the template. Computer-assisted analysis revealed the putative open reading frame encoded a protein of 323 amino acids with a predicted molecular weight of 35kDa. Amino acid alignment of MMP with other proteins of viruses in the same genus showed that they were mostly homologious, with identity of 91-99%, of which a transmembrane domain was predicted. MMP fusion protein was deleted the transmembrane domain when prokaryotic expressing, and the antibodies against it was prepared. The intracellular distribution and western blot analysis indicated that the MMP gene coded a membrane protein.
Document Type学位论文
Recommended Citation
GB/T 7714
徐维. 蛙虹彩病毒RGV一个膜蛋白基因的鉴定[D]. 水生生物研究所. 中国科学院水生生物研究所,2007.
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