摘要
以花椰菜品种‘闽花60天’根尖为材料,采用微细玻璃针法随机分离了40多条花椰菜的单染色体,并用LA-PCR(linker-adaptorPCR)对分离的单染色体进行了体外扩增,研究了单染色体的Sau3A酶切时间对LA-PCR扩增片段大小的影响。结果表明,通过对单染色体的不完全酶切可以显著提高LA-PCR扩增片段的大小;Dot-blotting验证的结果表明,单染色体的扩增产物确实来自其基因组,但Dot-blotting不能有效地鉴别花椰菜的9条染色体。
More than 40 single-chromosomes of the cultivar Minhua-60-day, in cauliflower(Brassica oleracea L.var. botrytis L.) were randomly isolated with micro-glass-needles by means of micromanipulation. LA-PCR was carried out in isolated single chromosomes DNA. The relationship between the time of digestion with Sau 3A and the length of amplified LA-PCR fragments was also investigated. Longer fragments were obtained in case of incomplete digestion of single-chromosome with Sau 3A. The results of Dot-blotting hybridization indicated that the PCR products of single chromosome came from the genomic DNA, 40 isolated single chromosomes couldn’t be divided into 9 groups with Dot-blotting.
出处
《中国农业科学》
CAS
CSCD
北大核心
2004年第11期1760-1765,共6页
Scientia Agricultura Sinica
基金
福建省青年科技人才创新资助项目(2001J048)
