摘要
目的建立一种简便、快捷、准确的检测方法用于近交系小鼠的遗传检测。方法根据近交系小鼠的H-2基因序列设计相应的探针,并标记生物素,利用微孔板Southern杂交技术,使探针与模板DNA杂交,再加入亲和素标记的辣根过氧化物酶进行酶显色反应,通过酶标仪检测杂交结果,以确定近交系小鼠的基因型。结果57BL/6和C57BL/10为H-2b型;DBA/2和Scid为H-2d型;615和C3H为H-2k型;NCPC/2、TA1、TA2和T739均为H-2b型。结论通过Southern杂交检测可以确定近交系小鼠的基因型。该检测方法简便、易行,检测结果客观,可以应用于近交系小鼠的遗传检测。
Objective In order to improve the efficiency and sensitivity of genetic monitoring of inbred strain mice, a method of Southern blot was developed in this study. Method Approximately 30 bp probe was established and inserted into the gene fragment of H-2D and H-2K, respectively. The probes were biotin-labelled and the DNA was fixed on microplate, and the biotin-labelled probes were reacted with avidin-HRP. Using the Southern blot method and microplate reader, according to the results whether the tested DNA and the value of OD of the specific probe hole is more than or equal to 0.2, and whether the tested DNA and the value of OD of the specific probe hole/tbe negative DNA and the value of OD of the specific probe hole is more than or equal to 2.1, different H-2 genotypes of the inbred strain mice can be determined. Results The H-2 genotype of different inbred strain mice could be distinguished by reading the Southern blot products. Conclusion The Sourtbern blot technique developed in this study can be successfully used in monitoring the H-2 gene and overcome the shortages of conventional method. Compared with previous methods, the newly developed test method is fast, sensitive and relaible, and simple to use.
出处
《中国实验动物学报》
CAS
CSCD
2008年第1期62-64,共3页
Acta Laboratorium Animalis Scientia Sinica
作者简介
马丽颖（1972-），女，助理研究员，研究方向：遗传检测。E-mail：maliyinglw@sina．com。
