Tian X, Chen L, Fang F
Cardiovascular Institute, CAMS and PUMC.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1994 Aug;16(4):302-8.
A 24kb fragment containing full-length renin-2 gene and its flanking sequence was obtained by digesting the pKG-R2D with XhoI. This fragment was cleaved with single enzyme (EcoRI, KpnI and BamHI), and combined enzymes (EcoRI/KpnI, KpnI/BamHI and BamHI/EcoRI), respectively. The digests were electrophoresed on 0.8% agarose plates and transferred onto NC membranes. Radioactive probes of 735bp and 1400bp, templates of which were from half and full-length of renin-1 cDNA, respectively were used for Southern hybridization. On the basis of electrophoresis and hybridization patterns, a restriction map was constructed in this paper.
用XhoI酶切pKG-R2D,获得了一个包含肾素-2基因全长及其侧翼序列的24kb片段。该片段分别用单酶(EcoRI、KpnI和BamHI)以及复合酶(EcoRI/KpnI、KpnI/BamHI和BamHI/EcoRI)进行切割。酶切产物在0.8%琼脂糖平板上进行电泳,然后转移到硝酸纤维素膜上。分别以肾素-1 cDNA的一半和全长为模板的735bp和1400bp放射性探针用于Southern杂交。根据电泳和杂交图谱,本文构建了一个限制性酶切图谱。