作　　者： ; ; ; ;

机构地区： 深圳大学生命科学学院

出　　处： 《生物技术通报》 2013年第4期85-89,共5页

摘　　要： 以莱茵衣藻CC-849为材料，提取基因组DNA，利用BamHⅠ和Bgl Ⅱ对基因组DNA进行酶解，获得了可用于构建基因组文库的6—12kb的基因组片段，并浓缩至200ng／pL。该片段与λDNA载体连接，经噬菌体蛋白包装、侵染大肠杆菌XL1-blue后，获得了莱茵衣藻基因组文库。该文库的滴度为2．12×10^5pfu／mL，共有转化子4．26×10^4个，插入片段的平均长度约为9kb，扩增后基因组文库滴度为9．5×10^6pfu／mL。 The genomic DNA isolated from Chlamydomonas reinhardtii CC-849 was digested with BamHⅠ and Bgl Ⅱ. The 6-12 kb genomic DNA fragments were recovered from agarose gel and concentrated to 200 ng/μL. Then they were inserted into EDNA ZAP expression vector, packed by packaging extracts of λ phage and transformed into Escherichia coli XL1-Blue. Finally, the genomic DNA library was obtained with an average insert size of 9 kb. Results showed that the titer of genomic library was 2.12 × 10^5 pfu/mL and contained 4.26 × 10^4 clones. After amplified, the titer of genomic library was up to 9.5× 10^6 pfu/mL.

关 键 词： 莱茵衣藻 基因组文库 噬菌体 表达载体

领　　域： [生物学]