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目的:克隆莲藕CDPK基因的cDNA全长并对其进行序列分析。方法:根据已有的ESTs序列,设计3’端RACE引物,运用RACE技术克隆莲藕CDPK基因cDNA全长。结果:克隆得到一条含有poly(A)尾,长度为2 039bp的全长cDNA序列,包括374bp的3’非编码区和1 665bp的开放阅读框,编码554个氨基酸。其核苷酸序列与其他作物CDPKs的同源性为80%、80%、78%和71%。其氨基酸序列与NCBI数据库中葡萄CDPK、大豆CDPK、蓖麻CDPK及曼陀罗CDPK1的同源性较高,分别为84%、82%、83%和81%,因此把研究得到的基因命名为LrCDPK。结论:CDPK基因的分离,为进一步研究该基因在莲藕生长发育过程中的功能奠定基础。
OBJECTIVE: To clone the cDNA of CDPK gene from lotus root and sequence analysis. Methods: According to the existing ESTs sequences, a 3 ’RACE primer was designed and the cDNA of CDPK gene was cloned by RACE. Results: A full-length cDNA with a length of 2 039 bp was obtained by cloning. It contained 374 bp 3 ’non-coding region and 1 665 bp open reading frame and encoded 554 amino acids. Its nucleotide sequence has 80%, 80%, 78% and 71% identity with other crop CDPKs. Its amino acid sequence is 84%, 82%, 83% and 81% homologous to the grape CDPK, soybean CDPK, ricin CDPK and mandarin CDPK1 in the NCBI database, so the gene was named as LrCDPK. Conclusion: The isolation of CDPK gene provides a basis for further study of the function of this gene in the growth and development of lotus root.