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本研究从陆地棉耐盐品种中9 409中克隆得到线粒体基因rps12(ribosomal protein S12,30S核糖体蛋白S12),并对该基因进行生物信息学、实时荧光定量PCR和原核表达分析。结果表明:陆地棉线粒体基因rps12的ORF全长为372 bp,编码123个氨基酸,分子量约为14.102 k D,等点电为11.11。rps12基因的表达具有组织特异性,且受盐胁迫诱导。另外,在大肠杆菌BL21中,重组原核表达质粒p ET28a-rps12可经IPTG诱导表达。研究结果为今后进一步研究陆地棉线粒体基因rps12的功能和应用奠定了分子生物学基础。
In this study, the mitochondrial gene rps12 (ribosomal protein S12, 30S ribosomal protein S12) was cloned from 9 409 of upland cotton cultivars. The gene was analyzed by bioinformatics, real-time fluorescence quantitative PCR and prokaryotic expression. The results showed that the ORF of mitochondrial rps12 gene was up to 372 bp in length and encoded 123 amino acids with a molecular weight of about 14.102 kD and an isoelectric point of 11.11. The expression of rps12 gene is tissue-specific and is induced by salt stress. In addition, in E. coli BL21, the recombinant prokaryotic expression plasmid pET28a-rps12 can be induced by IPTG. The results laid the foundation of molecular biology for further study on the function and application of mitochondrial rps12 gene in upland cotton.