文献类型：期刊文献

中文题名：千年桐SAD基因克隆与分析及其丝状真菌表达载体构建

英文题名：Clonging and Characterization of SAD from Vernicia montana and Construction of Filamentous Fungi Expression Vector

作者：范妙华[1] 李纪元[1] 范正琪[1] 田敏[1] 倪穗[2]

第一作者：范妙华

机构：[1]中国林业科学研究院亚热带林业研究所,浙江富阳311400;[2]宁波大学,浙江宁波315211

年份：2008

卷号：28

期号：1

起止页码：18-22

中文期刊名：西北植物学报

外文期刊名：Acta Botanica Boreali-Occidentalia Sinica

收录：CSTPCD;;北大核心:【北大核心2004】；CSCD:【CSCD2011_2012】；

基金：浙江省重大项目(2006C12030;2005C12003)

语种：中文

中文关键词：千年桐;硬脂酰脱饱和酶;丝状真菌;表达载体

外文关键词：Vernicia montana ; stearoyl-ACP desaturase（SAD） ; filamentous fungi ; expression vector

分类号：Q789

摘要：以发育中的千年桐种子总RNA为模板,通过RT-PCR方法扩增得到硬脂酰脱饱和酶基因SAD的cDNA序列。该序列长度为1 191 bp,编码396个氨基酸。推测的分子量为45 541.01 u,等电点pI为6.05。BLAST分析表明,该cDNA序列与其它已登录的SAD基因cDNA序列一致性最高可达93.1%;编码的氨基酸蛋白序列性一致最高为89%。同时,构建了由构巢曲霉3-磷酸甘油醛脱氢酶基因的gpdA启动子驱动的丝状真菌表达载体,通过冻融法转入农杆菌中,PCR鉴定表明,pBAR-SAD已转入农杆菌EHA105中,成功构建了农杆菌工程菌株。
The total RNA was extracted from the developing seeds of Vernicia montana, and the sequence of SAD was obtained by RT-PCR reaction,which was 1 191 bp long and encoded a protein of 396 amino acides with Mr=45 541.01 u and pI=6.05. The deduced nucleotides had 93.1% identity in comparison with the reported datas and 89% identity with other amino acids. A filamentous fungi expression vector,which was controlled by promoter of Aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase gene, was con- structed. The recombinant plasmid DNA was transformed into Agrobacterium tumefaciens by the FreezeThaw Methnd It determined that nBAR-SAD has been transformed into EHA105 by PCR.