Zhang Peng, Chen Yue-Qin, Liu Yi-Fei, Zhou Hui, Qu Liang-Hu
Key Laboratory of Gene Engineering of the Ministry of Education, Biotechnology Research Center, Zhongshan University, Guangzhou 510275, People's Republic of China.
Gene. 2003 Jun 5;311:93-8. doi: 10.1016/s0378-1119(03)00560-2.
The mitochondrial genome of the Chinese giant salamander Andrias davidianus was isolated using the long-and-accurate polymerase chain reaction (LA PCR) method. The sequencing work adopted the shotgun strategy accompanying with seven internal primers to cover the gaps where overlapping clones were not available. The entire mtDNA sequence is 16,503 bp long, with a gene content of 13 protein-coding, two ribosomal RNA and 22 transfer RNA genes, and order identical to that observed in most other vertebrates except for an additional 318 bp non-coding sequence between tRNA-Thr and tRNA-Pro genes. In order to carry out molecular phylogenetic analyses, all 13 protein sequences deduced from whole mitochondrial genomes for eight vertebrate species (six amphibians, two lobe-finned fishes) were combined to a single data set. This data set was refined by the program Gblocks using a stringent parameter setting and then subjected to MP, ML and NJ analyses. Thus phylogenetic relationships among living amphibians were discussed.
采用长片段精确聚合酶链反应（LA PCR）方法分离了中国大鲵（Andrias davidianus）的线粒体基因组。测序工作采用鸟枪法策略并辅以七条内部引物，以覆盖重叠克隆无法覆盖的缺口。整个线粒体DNA序列长度为16,503 bp，基因组成包括13个蛋白质编码基因、两个核糖体RNA基因和22个转运RNA基因，其排列顺序与大多数其他脊椎动物相同，只是在tRNA-Thr和tRNA-Pro基因之间有一个额外的318 bp非编码序列。为了进行分子系统发育分析，将八个脊椎动物物种（六种两栖动物、两种肉鳍鱼类）的整个线粒体基因组推导的所有13个蛋白质序列合并为一个单一数据集。该数据集通过程序Gblocks使用严格的参数设置进行优化，然后进行MP、ML和NJ分析。从而讨论了现存两栖动物之间的系统发育关系。
