|
|||||
|
|
| SSCP和HMA方法在马MHC-Ⅰ类分子基因多态性研究中的应用 | |
| 引用本文： | 项伟,马建,王雪峰,赵玉军,周建华.SSCP和HMA方法在马MHC-Ⅰ类分子基因多态性研究中的应用[J].遗传,2008,30(12):1635-1639. |
| 作者姓名： | 项伟 马建 王雪峰 赵玉军 周建华 |
| 作者单位： | [1]沈阳农业大学畜牧兽医学院,沈阳110161 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨150001 [3]东北林业大学野生动物资源学院,哈尔滨150040 [4]内蒙古农业大学动物科学与医学学院,呼和浩特010018 |
| 基金项目： | 国家自然科学基金(项目编号：30771994),黑龙江省发展高新技术产业专项资金(项目编号：FW05B007) |
| 摘 要： | 摘要: 文章使用SSCP和HMA两种基于聚丙烯酰胺凝胶电泳的方法对马MHC-I类分子基因多态性进行了分析。在应用SSCP法进行分析时, 尽管经过实验条件优化, 仍未得到对MHC-I基因理想的分离效果, 提示该方法对分离多态性较高的基因有一定局限性。在对HMA法用参考标准DNA对影响DNA分子构象的温度和变性剂浓度等实验条件进行优化后, 获得了对马MHC-I类分子基因较好的分离效果。6、7、8、9和10号马的样本在相对应泳道上分别出现了6、5、6、5和7个条带。从凝胶中进行DNA条带回收后克隆测序的结果表明, 这一方法可以有效地分离高度多态性的MHC-I类分子基因。 |
| 关 键 词： | SSCP HMA 基因多态性 MHC-I类分子基因 |
| 收稿时间： | 2008-03-12 |
| 修稿时间： | 2008-04-21 |
Applications of SSCP and HMA for polymorphic analysis of horse MHC-I alleles |
|
| Institution: | 1. Institutes of Animal Husbandry and Veterinary Medicine, Shenyang Agricultural University, Shenyang 100161, China; 2. National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China; 3. College of Wildlife Resources, Northeast Forestry University, Harbin 150040, China; 4. College of Animal Science and Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China |
| Abstract: | Abstract: In this article, we report the analysis of genetic polymorphisms of horse MHC-I molecules by SSCP and HMA, which are methods based on the technique of polyacrylamide gel electrophoresis (PAGE). Our results showed that SSCP was not a suitable method for the analysis of genetic polymorphisms of horse MHC-I molecules due to the failure in gener-ating satisfied separation of DNA fragments, even if experimental conditions were optimized. However, the HMA method produced clearly separated DNA fragments of horse MHC-I molecules, after the experimental conditions, such as the run-ning temperature and the concentration of detergent, were optimized by using a reference plasmid. PCR-amplified samples from horses No. 6, No. 7, No. 8, No. 9 and No. 10 generated 6, 5, 6, 5, and 7 bands, respectively, in corresponding lanes of the polyacrylamide gel. DNA fragments in each band cut from the gel were amplified by PCR using a second pair of prim-ers, and were cloned for sequencing. Alignment analysis of these sequences revealed that HMA was a proper method to efficiently analyze the polymorphisms of MHC-I molecule genes. |
| Keywords: | |
| 本文献已被 维普 等数据库收录！ | |
| 点击此处可从《遗传》浏览原始摘要信息 | |
| 点击此处可从《遗传》下载免费的PDF全文 | |