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为了开发沙地柏(Sabina vulgaris)的逆转座子基因资源,分析其在沙地柏基因组中的多样性和系统进化关系,采用兼并-PCR技术克隆了22条Ty1-copia类逆转座子逆转录酶序列。所克隆的核苷酸序列除去两端引物后的长度范围为239~247bp,同源性范围为49.6%~98.4%,显示出较高的异质性。核苷酸聚类分析结果显示,该22条序列由5个家族组成;氨基酸序列分析结果显示,有7条序列出现终止密码子突变,4条序列表现移框突变;且将这些逆转录酶的氨基酸序列与其它物种同一类型逆转录酶的氨基酸序列进行聚类分析表明,它们可能与其它物种的Ty1-copia逆转录酶有共同的起源。以上研究结果证实了沙地柏基因组中逆转座子的多样性,可为进一步利用逆转座子进行沙地柏的品种鉴定和遗传多样性分析奠定基础。
In order to develop the retrotransposon gene resources of Sabina vulgaris and analyze its diversity and phylogenetic relationship in Sabina vulgaris, 22 Ty1-copia retrotransposons were cloned using annex-PCR technique Enzyme sequence. The length of the cloned nucleotide sequence was 239 ~ 247 bp after removing the two end primers, and the homology ranged from 49.6% to 98.4%, indicating high heterogeneity. Nucleotide cluster analysis showed that the 22 sequences consisted of 5 families. Amino acid sequence analysis showed that there were 7 stop codons in the sequence and 4 sequences in the frame. Cluster analysis of the amino acid sequences with the amino acid sequences of the same type of reverse transcriptase from other species suggests that they may share a common origin with the Ty1-copia reverse transcriptase from other species. The above results confirm the diversity of retrotransposons in Sabina vulgaris genome and lay the foundation for the further identification and genetic diversity analysis of Sabina vulgaris with retrotransposons.