[Amplification, clone and identification of the specific fragments of tumor metastasis-suppressor gene nm23-H1 and nm23-H2].
A series of DNA primers specific for the specific fragments of nm23-H1 and nm23-H2 were designed and synthesized. The specific fragments of nm23-H1(185 bp) and nm23-H2(145 bp) were amplified from human blood DNA by using polymerase chain reaction (PCR). The recovered PCR products were treated with Klenow fragment, inserted into pGEM-3zf(+) vector with blunt-end ligation, and then transformed into competent cell JM109. The positive colonies were directly identified by colour screening on indicator plates. The recombinant plasmids were digested by Alu I and identified by PCR. The results showed that the authors had obtained the specific fragments of nm23-H1 and nm23-H2 respectively, and these specific fragments could be used for study the expression of nm23-H1 and nm23-H2 seperately.
Li
C
C
West China University of Medical Sciences, China.
Lu
Y
Y
Wang
R
R
D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D005784:Gene Amplification; D016147:Genes, Tumor Suppressor; D006801:Humans; D020559:Monomeric GTP-Binding Proteins / Q000235:genetics*; D054778:NM23 Nucleoside Diphosphate Kinases; D009701:Nucleoside-Diphosphate Kinase; D016133:Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics*
D054778:NM23 Nucleoside Diphosphate Kinases; D014157:Transcription Factors; D004247:DNA; C517938:NME1 protein, human; D009701:Nucleoside-Diphosphate Kinase; D020559:Monomeric GTP-Binding Proteins