谢瓦氏曲霉间型变种ychf 基因cdna 全长克隆及序列分析cloning .pdf

基因组学与应用生物学，2013 年，第32 卷，第2 期，第185-189 页 Genomics and Applied Biology, 2013, Vol.32, No.2, 185-189 研究报告 Research Report 谢瓦氏曲霉间型变种YchF 基因cDNA 全长克隆及序列分析 1,2 2,3 2,3 2,4* 李孝霞 谭玉梅 刘永翔 刘作易 1 贵州大学, 贵阳, 550025; 2 贵州省农业生物技术重点实验室, 贵阳, 550006; 3 贵州省生物技术研究所, 贵阳, 550006; 4 贵州省农业科学院, 贵 阳, 550006 * 通讯作者, liuzuoyi@ 摘 要 从谢瓦氏曲霉间型变种抑制性差减杂交(SSH)文库中，筛选到一个子囊孢子时期差异表达的序列标 签A0128.860 ，以其序列为模板设计特异性引物，利用RT-PCR 及RACE 技术克隆获得了该基因的全长cD- NA ，命名为eYchF 。生物信息学分析显示，该基因包含一个1 182 bp 的完整开放阅读框(ORF)，编码393 个氨 基酸，相对分子质量为43.466 9 kD ，预测的理论等电点为6.99 ，包括一个保守的G domain (guanine nu- cleotide-binding domain)和一个TGS (ThrRS, GTPase, SpoT) domain ，为疏水蛋白。序列同源性分析表明：该基 因与棒曲霉(Aspergillus clavatus NRRL 1) 的GTP 结合蛋白YchF 相似度为84%，谢瓦氏曲霉间型变种YchF 基因的克隆及生物信息学分析为YchF 的进一步研究奠定了基础。 关键词 谢瓦氏曲霉间型变种, YchF 基因, RACE Cloning and Sequence Analysis of the Full Length YchF cDNA in Asper - gillus chevalieri var. intermedius Li Xiaoxia 1,2 Tan Yumei 2,3 Liu Yongxiang 2,3 Liu Zuoyi 2,4* 1 Guizhou University, Guiyang, 550025; 2 Guizhou Key Laboratory for Agricultural Biotechnology, Guiyang, 550006; 3 Guizhou Institute of Biotech- nology, Guiyang, 550006; 4 Guizhou Academy of Agricultural Science, Guiyang, 550006 * Corresponding author, liuzuoyi@ DOI: 10.3969/gab.032.000185 Abstract TAn expression sequence tag (EST), A0128.860, which is differentially expressed in the period of ascospore, was selected from the Suppression Subtractive Hybridization (SSH) library. The complete cDNA of eYchF gene was cloned by RT-PCR and RACE with primers designed based on the sequence of A0128.860. Anal- ysis of bioinformatics showed that the gene contains a uninterrupted open reading frame of 1 182 bp and encoded a hydrophobic protein of 393 amino acid residues containing a G domain and a TGS domain. The estimated molecular weight and isoelectric point of the putative protein were 43.466 9 kD and 6.99, respectively. Analysis of the nucleotide sequen