| 哈茨木霉天冬氨酸蛋白酶基因SA76的3＇RACE扩增和序列分析 |
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| 引用本文： | 刘燕,杨谦.哈茨木霉天冬氨酸蛋白酶基因SA76的3＇RACE扩增和序列分析[J].林业研究,2007,18(2):139-143. |
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| 作者姓名： | 刘燕 杨谦 |
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| 作者单位： | 哈尔滨工业大学生命科学系 哈尔滨150001 |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | 由EST获得全长cDNA对于结构基因组学和功能基因组学都是至关重要的,cDNA末端快速扩增技术RACE是该领域中的重要研究方法.利用BD SMART RACE技术扩增编码分泌天冬氨酸蛋白酶SA76基因的3＇末端,将其与哈茨木霉cDNA文库中的SA76基因的EST序列进行序列拼接,获得2019bp的全长cDNA序列,其开放读码框长1593bp,5＇非编码区266bp,3＇非编码区201bp,编码530个氨基酸,有信号肽.哈茨木霉天冬氨酸蛋白酶基因与玉蜀黍赤霉、粗糙脉孢菌、球毛壳菌天冬氨酸蛋白酶基因的同源性分别为53%, 37%, 36%.利用BD SMART RACE技术首次从哈茨木霉中克隆天冬氨酸蛋白酶基因,为验证SA76基因的功能奠定基础,为进一步研究蛋白酶的作用机制及生物防治功能提供依据.
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| 关 键 词： | 哈茨木霉 天冬氨酸蛋白酶 SA76 基因扩增 序列分析 |
| 文章编号： | 1007-662X（2007）02-0139-05 |
| 收稿时间： | 23 December 2006 |
| 修稿时间： | 2006-12-232007-02-26 |
The RACE amplifying and sequence analyzing of secreted aspartic proteinase gene SA76 of <Emphasis Type="Italic">Trichoderma harzianum</Emphasis> |
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| Liu?Yan,Yang?Qian.The RACE amplifying and sequence analyzing of secreted aspartic proteinase gene SA76 of Trichoderma harzianum[J].Journal of Forestry Research,2007,18(2):139-143. |
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| Authors: | Liu Yan Yang Qian |
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| Institution: | Department of Life Science and Engineering, Harbin Institute of Technology, Harbin 150001, P.R.China;Department of Life Science and Engineering, Harbin Institute of Technology, Harbin 150001, P.R.China |
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| Abstract: | Total RNA was isolated from mycelium of T. harzianum by Total RNA extraction kit, and two clear bands of rRNA (28S and 18S) were observed in agarose electrophoresis. By joining
the 3′end sequence with the known SA76 EST from cDNA library of T. harzianum, a full-length cDNA sequence of 2019bp was obtained, whose open reading frame contained 1593bp, a stop codon TAA, a 5′untranslated
region (5′UTR) of 266bp, a 3′untranslated region (3′UTR) of 201bp, and poly (A) 29 encoded a protein of 530 amino acids, had
a signal peptide. T. harzianum shared 53% identity of secreted aspartic proteinase gene with G. zeae, 37% with N. crassa and 36% with C. globosum. The full-length cDNA sequence of secreted aspartic proteinase gene from T. harzianum was cloned for the first time by using BD SMART RACE technique, which provides a foundation to obtain and validate functional
genes of T. harzianum.
Foundation project: This paper was supported by Development Program (863) of China (No. 2003AA241140)
Electronic supplementary material is available in the online version of this article at http://dxdoi.org/10.1007/s11676-007-0028-6
Biography: LIU Yan (1976–), female, Ph.D Candidate, in the department of Life Science, Harbin Industrial University, Harbin
150001, P. R. China. |
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| Keywords: | Trichoderma harzianum BD SMART RACE BD SMART RACE T harzianum secreted aspartic proteinase gene |
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