洞庭湖日本血吸虫疫区东方田鼠生化与分子遗传学标记的初步研究  

Studies on Biochemical and Molecular Genetic Markers of Microtus Fortis in Dongtinghu Lake Region of China

作　　者：胡维新[1] 杨榕[1] 许冰[1] 曾海涛[1] 刘新发[1] 彭兴华[1] 

机构地区：[1]中南大学湘雅医学院分子生物学研究中心,长沙410078

出　　处：《中国实验动物学报》2002年第4期205-209,共5页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家科技部"九.五"国家科技攻关项目 (专题合同编号 :96 A2 3 0 6 0 4);湖南省科技厅"九.五"科技攻关配套基金;国家自然科学基金 (批准号 :3 0 0 70 40 3 )资助。

摘　　要：目的　建立东方田鼠生化与分子遗传学标记检测法。方法　参考近交系小鼠、大鼠生化遗传标记检测方法 ,对东方田鼠某些同功酶进行活性测定。根据东方田鼠特异DNA序列 ,合成引物 ,扩增东方田鼠基因组DNA ,将PCR产物回收、序列分析并进行生物信息学分析。结果　东方田鼠Trf、Es 3和Ce 2三个生化位点呈现遗传多态性 ,而Id1、Mod 1、Car 2、Gpd 1、Gpi 1、Hbb、Es 1七个生化位点无遗传多态性。用合成的特异引物扩增东方田鼠基因组DNA ,得到了丰富的多态性资料。对其中的 2 0只东方田鼠的扩增产物进行测序并进行多序列比较 ,发现10个等位基因以及 16个单核苷酸多态性 (SNP)位点。结论　初步建立了东方田鼠生化及分子遗传学标记。分子遗传学标记与生化遗传标记相比 ,具有杂合率高、重复性好、易于操作等优点。这些结果为深入研究东方田鼠的遗传背景、生物进化规律积累了基础资料。ObjectiveTo set up preliminarily the biochemical and molecular genetic markers of Microtus Fortis in Dongtinghu Lake region of China. Methods According to the methods of biochemical genetic markers for inbred strains of mice and rats, \%the state quality standard for lab animals in China: genetics\% published by Bureau of China State Technique for Quality Control, some isoenzyme activities of Microtus Fortis were detected. The primers were synthesized on the basis of specific DNA fragment published before and the genomic DNA of Microtus Fortis was amplified. The PCR products were recovered and cloned, the DNA sequencing as well as bioinformatics analysis were performed. Results Ten biochemical genetic sites, that is, Id1, Mod -1, Car -2, Gpd -1, Gpi -1, Hbb, Es -1, Trf, Es -3 and Ce -2, for more than 50 Microtus Fortis were analyzed and compared with those of mice. The genetic frequencies were calculated for closed colony of Microtus Fortis. The biochemical genetic polymorphisms for Trf, Es -3 and Ce -2 were found in ten sites above. Genomic DNA of 41 Microtus Fortis was amplified by two slightly different specific pairs of primers according to the specific DNA fragment of Microtus Fortis. A large number of genetic polymorphisms in molecular genetics were found. After comparing the sequencing results of the specific DNA fragment of 20 Microtus Fortis, we got 10 alleles and found 16 genetic polymorphism sites in specific sequence of Microtus Fortis. The specific DNA fragment of Microtus Fortis has abundant single nucleotide polymorphisms (SNP). ConclusionsThe abundant genetic polymorphisms of biochemical and molecular genetic markers will provide basic materials for the genetic monitoring of closed colony and inbred strain of Microtus Fortis and will help us know more about the differences between different species and different individuals of Microtus Fortis.

关 键 词：洞庭湖 日本血吸虫疫区 东方田鼠 分子遗传学标记 生化遗传标记 抗感染性 单核苷酸多态性 

分 类 号：R383.24[医药卫生—医学寄生虫学] Q959[医药卫生—基础医学]