【作者】 刘晓冬；

【导师】 刘宝；

【作者基本信息】 东北师范大学 ， 细胞生物学， 2005， 硕士

【摘要】 利用 Ty1/copia-like 类反转座子反转座酶的保守位点设计引物,从人参和西洋参基因组中扩增得到长度为 240bp 左右的 PCR 产物。对扩增产物进行分离和克隆,并随机挑选了 80 多个克隆进行测序,结果得到了 60 个不同的序列。Blast X 分析表明,这些克隆均为真实的反转录酶序列。根据序列比对,这些序列可分为 7 组以及单个的 3 组。这些核苷酸序列组内和组间均具有较大的差异,说明其核酸序列的高度异质性。这些序列中有 47 个是完整的,其余的有移码突变、终止子或二者兼有。Southern 杂交结果显示了这些序列在人参和西洋参中的分布情况。甲基化分析表明这些序列在人参和西洋参中均被高度甲基化。更多还原

【Abstract】 Ty1-copia-like sequences were amplified from two relative Panax species (Panax ginseng and Panax quinquefolius) genomic DNA, by PCR with degenerate oligonucleotide primers corresponding to highly conserved domains in the Ty1-copia-like retrotransposons. PCR fragments of roughly 246bp were isolated and cloned, and 83 clones were obtained. According to alignment and phylogenetic analysis, all sequences were divided into ten groups. Among these families, seven ones were enclosed in phylogenetic tree and three ones had only one sequence. The nucleotide sequence similarity shown by computer analysis between inter- and intra-group was in a wide range, which indicates high level of sequence heterogeneity among these clones and groups. Among these sequences, 47 clones had unique nucleotide sequences (intact sequences), and the others had a frameshift, a stop condon, or both. Southern analysis revealed distribution of these sequences in two panax species. Methylation analysis showed there were high level of methylation in two panax species更多还原