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马铃薯天冬氨酸蛋白酶抑制剂基因克隆和结构分析
引用本文:王永胜,刘中大.马铃薯天冬氨酸蛋白酶抑制剂基因克隆和结构分析[J].内蒙古大学学报(自然科学版),1996,27(5):670-674.
作者姓名:王永胜  刘中大
作者单位:内蒙古大学生物学系
基金项目:国家自然科学基金,内蒙古自然科学基金
摘    要:以马铃薯基因组DNA为模板并基于天冬氨酸蛋白酶抑制剂基因的cDNA序列所设计的两个引物,通过PCR扩增得到666bp的天冬氨酸蛋白酶抑制剂基因编码区,并将此片段克隆到pUC18的SmaI位点.序列分析结果表明该基因除去末端一终止密码子外其可译框架编码221个氨基酸残基,前导肽包括32个氨基酸残基,故该抑制剂的成熟蛋白由189个氨基酸组成,第67位的精氨酸为抑制胰蛋白酶的活性中心.与cDNA相比核苷酸的同源性为94.3%,氨基酸的同源性为90%.基因DNA无内含子.

关 键 词:天冬氨酸蛋白酶抑制剂基因,基因结构,PCR

Primary Structure Analysis of Gene Encoding an Aspartic Proteinase Inhibitor from Potato
Wang Yongsheng,Liu Zhongda,Shi Yixin,Hu Tingmao.Primary Structure Analysis of Gene Encoding an Aspartic Proteinase Inhibitor from Potato[J].Acta Scientiarum Naturalium Universitatis Neimongol,1996,27(5):670-674.
Authors:Wang Yongsheng  Liu Zhongda  Shi Yixin  Hu Tingmao
Abstract:On the basis of the nucleic acid sequence of cDNA encoding an aspartic proteinase inhibitor,two primer were synthesized. Using the potato genomic DNA as a template,an aspartic proteinase inhibitor gene was synthesized by PCR. The gene was inserted into the SmaI site of pUC18 for constructing pAPI. The results of dot hybridization and sequence analysis showed that the pAPI contains the region encoding an aspartic proteinase inhibitor. An open reading frame of the gene encodes a precursor protein of 221 amino acids. The genomic DNA sequence shares 94. 3% nucleotide and 90% amino acid identity with cDNA pudlished before. There are no nitron sequences interupting the coding region.
Keywords:aspartic proteinase inhibitor gene  gene structure  PCR  
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