摘要
设计以 aad A基因置换衣藻 chl L基因并作为转基因衣藻筛选标记 ,将 chl L基因两侧的基因片段 clp P- trn L- pet B- chl L5′- 3′作为同源片段 ,构建衣藻叶绿体同源重组质粒 p64D.通过改进的基因枪法转化程序 ,将此质粒轰击入衣藻叶绿体 .经抗性筛选、暗培养及 PCR、Southern- blotting等鉴定 ,证实衣藻叶绿体基因组中大部分 chl L结构基因已被外源基因 aad
The recombinant plasmid p64D which contained the aad A gene as resistant screening marker to Chlamydomonas transformants and clp P trn L pet B chlL 5′ 3′ as homologous fragment was constructed and transferred to the chloroplast of C.reinhardtii by an improved biolistic method.The transformants were identified by PCR,Southern blotting after selecting in resistant medium and incubating in the dark.The results showed that the great part of the chlL structure genes in the chloroplast genome were replaced by aad A gene.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
2000年第1期72-76,共5页
Chinese Journal of Biochemistry and Molecular Biology
关键词
衣藻叶绿体
同源重组
chlL基因
同质化鉴定
Chlamydomonas chloroplast,Homologous recombination, chlL gene, aad A gene
