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[Cloning of mouse polycystic kidney disease 1 gene]
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 mouse 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.