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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

Primary Structure Analysis of Gene Encoding an Aspartic Proteinase Inhibitor from Potato
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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. 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.
出处 《内蒙古大学学报(自然科学版)》 CAS CSCD 1996年第5期670-674,共5页 Journal of Inner Mongolia University:Natural Science Edition
基金 国家自然科学基金 内蒙古自然科学基金
关键词 抗虫性育种 天冬氨酸 蛋白酶抑制剂 基因结构 aspartic proteinase inhibitor gene gene structure PCR
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