火龙果类反转录转座子反转录酶序列的克隆和分析.pdfVIP

园 艺 学 报 2012 ，39 （2 ）：265 –272 http: // www. ahs. ac. cn Acta Horticulturae Sinica E-mail: yuanyixuebao@126.com 火龙果 Ty1-copia 类反转录转座子反转录酶序列 的克隆及分析 范付华，乔 光，郑思成，文晓鹏* （贵州大学贵州省农业生物工程重点实验室，贵阳 550025 ） 摘 要：根据 Ty1-copia 类反转录转座子反转录酶的保守区设计简并引物，通过 PCR 扩增，从火龙 果基因组中扩增出 260 bp 左右的目标条带，经回收、克隆、测序及序列分析，获得了 23 条来源于火龙果 试管苗变异和非变异材料的反转录酶序列。通过系统演化分析，所得序列可分为 5 类，存在较高的异质 性；与母株相比，变异植株的反转录酶序列存在缺失、移码和终止密码子突变。经反转录酶氨基酸序列 对比，火龙果与其它物种间具有较高的同源性，表明在不同物种间可能存在反转录转座子的横向传递作 用。 关键词：火龙果；变异；Ty1-copia 类反转录转座子；反转录酶；异质性 中图分类号：S 668.9 文献标识码：A 文章编号：0513-353X （2012 ）02-0265-08 Cloning and Analysis of Reverse Transcriptase of Ty1-copia Retrotrans- posons in Hylocereus undatus * FAN Fu-hua ，QIAO Guang，ZHENG Si-cheng ，and WEN Xiao-peng （Guizhou Key Laboratory of Agricultural Bioengineering，Guizhou University，Guiyang 550025 ，China ） Abstract ：Using degenerate oligonucleotide primers corresponding to the conserved domains of the Ty1-copia retrotransposon reverse transcriptase ，a fragment of 260 bp was amplified by polymerase chain reaction （PCR ）from genomic DNA of Hylocereus undatus . The amplicons were recovered and cloned， then sequenced and analyzed. Twenty-three DNA sequences of reverse transcriptase were obtained from normal or somaclonal variant of Hylocereus undatus in vitro plants. The sequences can be classified into five groups with high heterogeneity through phylogenic analysis. The DNA sequences displayed mutation including deletion，frameshift or stop codon. A phylogenic tree was constructed based on the amino acid sequences from other species，indicating that they might share the common origin and that horizontal transmission of retrotransposons has occurred among the plants in the past.