Valent A, Meddeb M, Danglot G, Duverger A, Nguyen V C, Bernheim A
Laboratoire de Cytogénétique Oncologiques, UA 1967 CNRS, Institut Gustave-Roussy, Villejuif, France.
Hum Genet. 1996 Jul;98(1):12-5. doi: 10.1007/s004390050152.
Reverse transcriptase-polymerase chain reactions using foetal brain RNA with reverse and forward primers of the first, second and third NTRK4 region allowed us to obtain three amplified NTRK4 fragments. The specificity of amplified fragments was checked by digestion with restriction endonucleases AvrII, HindIII and PspII for the first, second and third regions, respectively. Each restriction site was specific for each amplified fragment. The fragment of the NTRK4 first region was also sequenced and the sequence determined was identical to the human NTRK4 sequence. The three amplified fragments were cloned in pBS. For the Southern technique, plasmid pBS-NTRK4a (with an insert of 1052 bp) detected a human 9-kb HindIII sequence which was localised unambiguously on chromosome 6. For fluorescence in situ hybridisation, the three plasmids, pBS-NTRK4a, pBS-NTRK4b (insert 924 bp) and pBS-NTRK4c (insert 1114 bp) were pooled and used as a probe. This NTRK4 probe was localised on 6p21. Of 50 metaphases analysed, 49 contained twin spot signals on both sister chromatids.
使用胎儿脑RNA以及神经营养酪氨酸激酶受体4（NTRK4）第一、第二和第三区域的正向和反向引物进行逆转录聚合酶链反应，使我们获得了三个扩增的NTRK4片段。分别用AvrII、HindIII和PspII限制性内切酶对第一、第二和第三区域的扩增片段进行消化，以检查扩增片段的特异性。每个限制性位点对每个扩增片段都是特异的。还对NTRK4第一区域的片段进行了测序，所确定的序列与人类NTRK4序列相同。将这三个扩增片段克隆到pBS载体中。对于Southern技术，质粒pBS-NTRK4a（插入片段为1052 bp）检测到一个人类9 kb的HindIII序列，该序列明确位于6号染色体上。对于荧光原位杂交，将三个质粒pBS-NTRK4a、pBS-NTRK4b（插入片段924 bp）和pBS-NTRK4c（插入片段1114 bp）混合用作探针。该NTRK4探针定位于6p21。在分析的50个中期相中，49个在两条姐妹染色单体上均含有双斑点信号。
