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【目的】从辣椒基因组中克隆Ty3-gypsy逆转录转座子逆转录酶序列,分析其家族特性及进化关系。【方法】根据Ty3-gypsy逆转录转座子逆转录酶保守区域设计简并引物,通过PCR扩增、回收、克隆、测序得到的目的基因序列。【结果】克隆从辣椒基因组14条逆转录转座子逆转录酶序列,序列大小均为340 bp左右。生物信息学分析表明,序列长度变化范围为310~345 bp,同源性范围为60.2%~99.1%。这些核苷酸序列具有较高的异质性,主要表现为终止密码子突变。聚类分析可将所得序列分为5大类,基于逆转录酶序列进行的不同物种进化树分析显示,第Ⅳ、Ⅴ类序列与荸荠、潘那利番茄等组成一大类,可能是逆转录转座子横向传递的结果。【结论】辣椒Ty3-gypsy逆转录转座子逆转录酶序列具有普遍性、异质性等特点,与荸荠、潘那利番茄等物种的同类型逆转座子可能有相同的起源。
【Objective】 The Ty3-gypsy retrotransposon reverse transcriptase sequence was cloned from pepper genome and its family characteristics and evolutionary relationships were analyzed. 【Method】 Degenerate primers were designed according to the conserved region of Ty3-gypsy retrotransposon reverse transcriptase. The target gene sequences were amplified, recovered, cloned and sequenced by PCR. 【Result】 The results showed that the sequence of 14 reverse transposon reverse transcriptases from pepper genome was about 340 bp. Bioinformatics analysis showed that the sequence length ranged from 310 bp to 345 bp, and the homology ranged from 60.2% to 99.1%. These nucleotide sequences have high heterogeneity and predominantly stop codon mutations. Cluster analysis can be divided into five categories, the phylogenetic tree analysis of different species based on reverse transcriptase sequence shows that the fourth and fifth sequences and water chestnuts, Panalpina tomato composition of a large category, may be reverse transcription Transposon results of the horizontal transfer. 【Conclusion】 The reverse transcription transposase sequence of Ty3-gypsy retrotransposon in peppers has the characteristics of universality and heterogeneity. It may have the same origin with the same type of retrotransposon in species such as water chestnut and Panalware.