摘要
目的 克隆小鼠的遗传型多囊肾病1 基因(polycystic kidney disease 1,PKD1),为产生PKD1 基因缺陷小鼠品系准备条件。方法 以正常小鼠基因组DNA 为模板、扩增小鼠PKD1 基因部分序列为探针,应用噬斑原位杂交技术筛选129SvTer小鼠基因组DNA 文库,并用Southern 杂交、亚克隆及测序等方法鉴定所得克隆。结果 筛选得到1 个阳性克隆,并证实了所得序列与基因库收录的序列一致。结论 已克隆到含有PKD1 基因目的区域的基因组DNA 片段,其结构符合构建目标载体的要求,为建立小鼠PKD1
Objective To obtain mouse polycystic kidney disease 1(PKD1) gene by plagues in situ hybridization from a genomic library.Methods With the use of partial mouse PKD1 genomic DNA fragments amplified by PCR as probes, a genomic library of 129SvTer muose in bacteriophage vector was screened by plagues in situ hybridization. The inserts of genomic DNA fragments were analyzed by Southern blot, subclone and verified by sequencing.Results A positive phage clone was obtained after four round screening. The sequence of the genomic DNA fragments inserted in the positive clone was in accordance with that of the Genebank.Conclusion The authors found that the length and structure of mouse PKD1 genomic DNA fragments obtained from a genomic library were available to construct a replacement targeting vector.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
1999年第6期364-367,共4页
Chinese Journal of Medical Genetics
基金
国家863 计划部分资助
关键词
多囊肾病
克隆
遗传型
PKD1
动物模型
Mouse polycystic kidney disease 1 gene Cloning Library screening
