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以雏菊为试验材料,采用PCR方法扩增雏菊rbcL基因并测序,并用DNAstar、PAML等生物软件对该基因进行生物信息学及其适应性进化研究。结果表明:雏菊rbcL基因序列全长1 508bp,编码区全长1 458bp,共编码485个氨基酸,与基因库中菊科其它属植物比较,核苷酸和氨基酸序列的同源性均超过了96%。rbcL蛋白二级结构主要有20个α螺旋、17个β折叠、33个转角以及一些无规则卷曲,适应性进化检测发现共有3个正选择氨基酸位点(94E,149Q,309M)。获得完整雏菊rbcL基因有助于更深地研究雏菊对其生境的适应,rbcL蛋白大亚基正选择位点的空间结构对于维持核酮糖结构具有重要的作用。
Daisy was used as experimental material to amplify and sequenced the daisy rbcL gene by PCR method. The bioinformatics and adaptive evolution of the gene were studied with DNAstar, PAML and other biological softwares. The results showed that the sequence of rbcL gene was 1 508 bp in length and 1 458 bp in length, encoding a total of 485 amino acids. Compared with other genera of Asteraceae in GenBank, the nucleotide and amino acid sequence identities exceeded 96 %. The secondary structure of rbcL protein consists of 20 α helices, 17 β-sheets, 33 turns and some random curls. Three positive selection amino acid sites (94E, 149Q, 309M) were identified by adaptive evolution assay. Obtaining the complete daisy rbcL gene will help to further understand the adaptation of daisy to its habitat. The spatial structure of the rbcL large subunit positive selection site plays an important role in maintaining ribulose structure.