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。
For exploiting retrotransposon gene resource,and analyzing diversity and phylogenetic evolution of retrotransposons in Sabina vulgaris genome,twenty-two of fragments encoding reverse transcriptase of Ty1-copia-like retrotransposon were cloned by degenerate-PCR.The length of the nucleotide sequences excluding primer varied from 239 to 247 bp,and homology ranged from 49.6% to 98.4%,showing high heterogeneity.Phylogenetic analysis of the nucleotide sequences showed that the twenty-two sequences consisted of five families.When they were translated into amino acids,stop codon mutation was found in seven sequences,and frameshift mutation was found in four sequences.The alignment analysis of amino acid sequences of Ty1-copia-like reverse transcriptases from S.vulgaris and other plant species suggested that they may have the same origin.These results demonstrated diversity of retrotransposon in S.vulgaris genome,and could prove to be useful for further study on genetic diversity and cultivar identification of S.vulgaris.
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