运用FISH技术将SSR标志定位于染色体上的研究  

The study of SSR mark localized on chromosome in FISH

作　　者：陆井元[1] 刘代[1] 吴尚红[1] 胡家金[1] 

机构地区：[1]湖南农业大学生物科学技术学院,湖南长沙410128

出　　处：《现代生物医学进展》2009年第12期2264-2266,2300,共4页Progress in Modern Biomedicine

基　　金：农业部"948"资助项目(2008-Z42);中国农科院蔬菜研究所提供序列及基金支持

摘　　要：目的:运用FISH将SSR标记定位于染色体上,并确定其具体属于哪条染色体。方法:从全基因组测序数据库中挑选SSR标记,再将该序列到Fosmid库中进行序列比对,得到末端序列与SSR序列相同的Fosmid,再利用该Fosmid制作荧光原位杂交的探针,将该探针运用FISH技术杂交到染色体上,同时结合Tpy1,Tpy3序列识别其具体位于哪条染色体上。结果:在荧光显微镜下可以直接观察到探针在染色体上的结合位点,利用相关拍摄软件就可以对其进行拍照。本实验得到了较好的FISH杂交图片,并结合Tpy1,Tpy3成功的将其定于黄瓜五号染色体上。结论:FISH技术可以让人直接观察到探针在染色体上的位置,运用此方法能将可用于进行分子遗传图谱整合的SSR标记准确定位于染色体上。本研究中,在Tpy1,Tpy3探针的帮助下,我们更直接确认了该标记位于黄瓜的第五号染色体上。这使该标记对黄瓜分子辅助育种与黄瓜分子遗传图谱的整合,可以提供直接而有用的帮助。Objective： To locate SSR mark on chromosome by FISH and to identify which chromosome it belongs to. Methods： We chose the SSR mark from a Gene bank and then we used this sequences blast in a Fosmid-bank and got a target Fosmid whose termi- nal sequence similar to the SSR sequence. We make this Fosmid into a probe of the FISH, at last, we localized this probe on the chromo- some by FISH. Combining the Tpyl and Tpy3 probe, we finally identified the exact chromosome that the probe belongs to. Results： We can see the direct localization of the probe on chromosomes under the fluorescence microscope, we use a Sensitive CCD camera （QIMAGING,RETIGA-SRV,FAST 1394） attached to an Olympus BX61 epifluorescence microscope（Tokyo,Japan）to catch these images. In this study,we got the hybridization images and we know it belonged to the fifth chromosome of cucumber. Conclusions： we can see the direct localization of the probe on chromosomes by FISH. In this technique we can locate the SSR mark which may be used for incorpo- rating different molecular genetic maps on chromosome. With the aid of Tpyl and Tpy3, we also identify this probe belongs to the fifth chromosome of cucumber. So this probe can offer a straight and useful aid in molecularaid breeding of cucumber and integeration in different molecular genetic maps.

关 键 词：FISH SSR 染色体 定位 

分 类 号：Q78[生物学—分子生物学]