摘要
从坛紫菜叶状体中提取基因组DNA,并用CTAB/NaCl纯化,得到质量较高的DNA,它无RNA污染,且OD260/OD280比值为1.89～1.95,适用于微卫星分子标记研究。根据坛紫菜微卫星DNA序列设计特异引物,进行坛紫菜微卫星的PCR扩增反应。用PopGen软件分析坛紫菜2个优良品系和1个野生种群间的Nei氏遗传相似系数和遗传距离分别在0.5760～0.6915和0.4251～0.6412,然后根据Nei氏遗传距离用MEGA软件构建了UPGMA聚类图,结果表明高蛋白和生长快这两个优良品系群体虽来自野生种群体,尽管表型差异不大,但从基因型来说它们已不同于野生种群体。同时还得到了3条可区分坛紫菜野生种群体及2个优良群体(生长快型、高蛋白型)的差异性条带:4#-117、25#-231、14#-302。
Genomic DNA was extracted from Porphyra haitanesis thallus according to the method described by Wattier et al.. After purification with CTAB and NaCl, high quality DNA could be obtained. The average size of genomic DNA was about 23 kb estimated by agarose gel electrophoresis which showed no RNA occurring on it, and the ratio of OD260/OD280 was around 1.89 - 1.95 determined spectrophotometrically, illustrating that the extracted DNA was good enough to meet the needs for analysis of microsatellite markers. The SSR- PCR amplification was performed using the special primers designed according to the microsatellite DNA sequences reported by Hu et al. in detail, and the PCR products were detected by polyacrylamide gel electrophoresis and stained by silver method. A Nei＇s genetic distance matrix could be constructed among three populations by using the PopGen32 software, demonstrating that Nei＇s genetic similarity was ranging between 0. 576 0 -0. 691 5 and Nei＇s genetic distance was 0. 425 1 -0.641 2. An UPGMA molecular dendrogram was constructed according to the Nei＇s genetic distance matrix by using the MEGA 4.0 software. It revealed that the two populations of fast-growing and high protein content first clustered, then clustered with wild population. Three special DNA bands named 25^# -231, 14^# -302 and 4^# -117 were also found among the wild type and the other two bred strains, namely fast-growing and high content of protein of this red seaweed.
出处
《上海海洋大学学报》
CAS
CSCD
北大核心
2009年第4期409-414,共6页
Journal of Shanghai Ocean University
基金
国家"八六三"高技术研究发展计划(2002AA603032
2006AA10A413)
上海市水产养殖重点学科建设项目资助(Y1101)
上海市高校E-研究院资助项目
关键词
坛紫菜
简单序列重复
微卫星DNA
Porphyra haitanensis
simple sequence repeat
microsatellite DNA
