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利用电子克隆的方法设计特异引物,克隆陆地棉的1个二硫键异构酶(Protein disulphide isomerase like)基因的全长序列。序列分析表明:该基因没有内含子,开放阅读框的长度为1 293bp,编码的多肽序列含有430个氨基酸。该基因与红麻HcPDIL5-2a基因具有97%的相似性,将该基因命名为棉花GhPDIL5-2a。取单核期花药进行半定量RT-PCR分析发现:GhPDIL5-2a在陆地棉洞A不育株和可育株花药中表达无差异,在供试的其他不同品种中的表达也无差异。
The specific primers were designed by electronic cloning method to clone the full length sequence of one disulfide isomerase like gene of G. hirsutum. Sequence analysis showed that the gene has no intron, the length of the open reading frame is 1 293bp, and the encoded polypeptide sequence contains 430 amino acids. The gene has a similarity of 97% with the kenaf HcPDIL5-2a gene, and the gene is named as cotton GhPDIL5-2a. Semi-quantitative RT-PCR analysis showed that GhPDIL5-2a had no difference in the Upland cotton A sterile and fertile anthers, and there was no difference among the other varieties tested.