辣椒果实发育相关基因的电子克隆及生物信息学分析

In Silico Cloning and Bioinformatics Analysis of Genes Related to Pepper Fruit Development

以cDNA-AFLP分析辣椒果实发育获得的长度为193 bp的TDF_(9-68-11)为研究对象,采用电子克隆、PCR验证和生物信息学分析等方法,研究了辣椒果实发育相关基因的电子克隆及生物信息学功能分析,以期为辣椒果实发育相关基因的研究提供参考依据。结果表明:经过电子克隆最终获得长为1 167 bp的拼接序列,设计引物PCR扩增和测序获得全长714 bp的序列,与拼接序列高度一致;该序列编码分子量为17.96 kD氨基酸链,其等电点为7.76偏中性;氨基酸链具有跨膜特性,可能存在5个磷酸化位点,该氨基酸链二级结构含有的α螺旋占42.3%、延伸链占19.6%、β转角占12.2%,以及26.4%的随机卷曲结构。氨基酸序列比对可以初步推断该氨基酸序列的功能与果实发育中醇脱氢相关生化反应有关。

The 193bp TDF9-68-11 obtained from cDNA-AFLP analysis of pepper fruit development was used as the research object.Electronic cloning,PCR validation,and bioinformatics analysis were used to study the electronic cloning and bioinformatics function analysis of genes related to pepper fruit development,in order to provide reference for the cloning and functional verification of genes related to pepper fruit development.The results showed that after in silico cloning,a sequence with a length of 1167bp was obtained.Primer PCR amplification and sequencing were designed to obtain a sequence with a total length of 714bp,which was highly consistent with the splicing sequence;The sequence encodes an amino acid chain with a molecular weight of 17.96kD and a neutral isoelectric point of 7.76;amino acids had transmembrane properties,and there might be 5phosphorylation sites in the amino acid sequence.The secondary structure of this amino acid containsα-spiral accounting for 42.3%,extended chain accounting for 19.6%,β-random curl structure with a corner of 12.2%and 26.4%.Amino acid sequence alignment could preliminarily infer that the function of this amino acid sequence was related to biochemical reactions related to alcohol dehydrogenation during fruit development.