作　　者： ; ; ;

机构地区： 深圳大学生命科学学院

出　　处： 《基因组学与应用生物学》 2013年第2期190-195,共6页

摘　　要： 本研究以雨生红球藻34—1n为材料，提取其基因组DNA，利用限制性内切酶Sau3AI对基因组DNA进行酶解，回收6～8kb的基因组DNA片段，并浓缩至200ng／i,zL。该片段与经BamHI酶切和去磷酸化处理后的pUC18载体连接，然后电击转化到受体菌Escherichia.coliDH5仅中，获得雨生红球藻34-1n的基因组文库。该文库的平均插入片段长度约为6．5kb，获得6x10s个克隆数。通过PCR筛选，由雨生红球藻基因组文库中获得含bktl序列的单克隆菌，与β-胡萝卜素氧化酶序列（GenBank：DQ086233．1）进行比对，结果表明bktl基因组序列含有6个外显子。本研究为进一步鉴定雨生红球藻相关基因提供了一个文库平台。 The genomic DNA extracted from Haematococcus pluvialis 34-1n was digested with Sau3A I. The genomic DNA fragments in length from 6 kb to 8 kb were recovered from agarose gel and concentrated to 200 ng/μL, which were inserted into pUC18 vector treated with BamH I and CIAP. The constructed plasmids were introduced into Escherichia. coli DH5ot by using electroporation. Finally, the genomic DNA library contained 6x105 clones was constructed with an average insert size of 6.5 kb. Clonies contained bktl gene were detemined by PCR technique from the constructed genomic DNA library. By comparing with the 13-C-4-oxygenase gene sequence （GenBank： DQ086233.1）, six exons were found in bktl genomic sequence. These study would provide a library platform to further identify noval genes from Haematococcus pluvialis.

关 键 词： 雨生红球藻 基因组文库 质粒载体 胡萝卜素酮化酶基因

领　　域： [生物学]