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为深入了解兴安落叶松的基因组构成及其逆转座子基因的特征,本研究克隆了其逆转座子逆转录酶序列,并进行了生物信息学分析。根据Tyl-copia型逆转座子逆转录酶的保守序列设计简并引物,并利用PCR技术获得了26条逆转座子逆转录酶序列。这些序列去除引物后的核苷酸长度范围为237~251 bp,序列间相似度为55.9%~90.9%,可被分为6个家族。氨基酸预测结果显示,有3条序列存在移框突变,9条序列存在终止密码子突变。系统进化分析显示,所克隆的17条没有发生终止密码子突变的序列分别与处于不同进化阶段的其他植物的Tyl-copia型逆转录酶具有共同的起源。研究结果证实兴安落叶松含有类型丰富的Tyl-copia型逆转座子,可为该物种逆转座子基因资源的开发和利用提供依据。
In order to further understand the genomic constitution of Larix gmelinii and the characteristics of its retrotransposon genes, this study cloned its reverse transcriptase sequence and carried out bioinformatics analysis. The degenerate primers were designed according to the conserved sequence of Tyl-copia retrotransposon reverse transcriptase, and 26 reverse transposase reverse transcriptase sequences were obtained by PCR. The deduced nucleotides of these sequences ranged from 237 to 251 bp in length with 55.9% to 90.9% similarity between the sequences and could be divided into 6 families. The results of amino acid prediction showed that there were three frameshift mutations and nine of them had stop codon mutations. Phylogenetic analysis showed that the cloned 17 sequences that did not undergo stop codon mutation shared common origin with Tyl-copia reverse transcriptase from other plants at different stages of evolution. The results of this study confirm that Larix gmelinii contains a large number of Tyl-copia retrotransposons, which may provide a basis for the development and utilization of the retrotransposon gene resources in this species.