成人型多囊肾病致病基因PKD1区新DNA片段的定位克隆

Positional cloning of a novel DNA fragment in the region of PKD1

目的为了分离成人型多囊肾病致病基因ＰＫＤ１区近侧端新的探针。方法采用定向跳跃克隆ＰＣＲ技术分离了一个ＤＮＡ片段，通过反向ＰＣＲ、缺失杂交定位，限制性内切酶谱分析以及ＤＮＡ序列分析，并与ＰＫＤ１区物理图谱及最新的ＤＮＡ数据库比较。结果证实该克隆片段是一个新的ＤＮＡ序列，位于ＰＫＤ１区ＮＫ９２．６ＳＨ１．０近侧的ＭｌｕⅠ位点处，距克隆起始位点ＡＪ１约４０ｋｂ。结论由于该位点位于ＰＫＤ１可能存在的另一候选基因的潜在区域内，故可作为进一步筛选ＰＫＤ１新候选基因的探针使用。

bjective To confirm the possibility that another candidate gene may exist likely mapped near the locus of NK926 SH10 in PKD1 region. Methods Using oriented jumping cloning PCR, a DNA fragment expected to be around the Mlu Ⅰ site, which is about 40kb distal from the jumping point (AJ1 locus) and 10kb proximal to NK 926 SH10 locus, has been cloned. Results Its genomic location expected was supported not only by the evidence from inverse PCR, but also by those from experiments of deletion hybridization and restriction mapping. Conclusion This cloned fragment could be used as a probe in selection of the new PKD1 candidate gene or other neighbouring genes in PKD1 region.