摘要
对拟南芥氰丙氨酸合酶(cyanoalanine synthase,CAS)基因进行了生物信息学分析,并构建了CAS合酶基因的超表达载体,以期为其后续功能研究奠定基础。生物信息学分析结果表明,编码拟南芥CAS合酶的CYS-C1和CYS-D1基因均含有10个外显子和9个内含子,定位于3号染色体,而CYS-D2基因有9个外显子和8个内含子,定位于5号染色体;3个基因编码的蛋白氨基酸序列相似度较高,CYS-C1蛋白偏碱性且主要在线粒体中起作用,而CYS-D1和CYS-D2蛋白偏酸性,主要在细胞质中起作用。通过RT-PCR扩增了拟南芥CYS-C1、CYS-D1和CYS-D2基因片段,并构建了超表达载体p BI121-35S-CYS-C1、p BI121-35S-CYS-D1和p BI121-35S-CYS-D2,经检测重组质粒已转化到农杆菌GV3101中。
The CAS genes were analyzed by bioinformatics methods and over-expression vectors of CAS genes were constructed. Bioinformatics analysis show that CYS-C1 and CYS-D1 have ten exons and nineintrons,while CYS-D2 has nine exons and eight intron. CYS-C1 and CYS-D1 locate at chromosome 3while CYS-D2 locates at chromosome 5. Furthermore,the amino acid sequences of CAS genes share ahigh similarity. However,CYS-C1 is basic protein and functions at mitochondria,whereas CYS-D1 and CYS- D2 are acidic protein and function at cytoplasm. The over- expression vectors of p BI121- 35SCYS- C1,p BI121- 35S- CYS- D1 and p BI121- 35S- CYS- D2 were constructed,and the recombinantplasmids were transformed into agrobacterium GV3101.
出处
《江汉大学学报（自然科学版）》
2015年第2期150-157,共8页
Journal of Jianghan University：Natural Science Edition
基金
国家自然科学基金项目(31400242)
武汉生物工程学院应用校本研究项目(2014K29)
