Research regarding oxygen-sensitive K channel existing in the ductus arteriosus.

猪动脉导管分离基因的建立：新生仔猪140头，于出生后即刻分离动脉导管组织、主动脉、肺动脉。从导管、主动脉和肺动脉中提取总RNA和聚A RNA。用RT-PCR方法从RNA中扩增出c DNA。将该基因克隆入载体，构建了文库。插入DNA的长度在1.7~2.7kb之间。用从大鼠、小鼠和人获得的Kv探针筛选猪动脉导管的cDNA文库。从动脉导管组织中克隆出Kv1、Kv2和Kv9.3基因。同源聚合酶链式反应筛选文库：以Kv基因的保守部分为模板设计引物，以其为模板进行聚合酶链式反应。比较导管、主动脉和肺动脉的PCR扩增产物。选择仅在导管中扩增的PCR产物进行克隆。克隆了在导管中表达的KV家族，它们分别属于1.5、2.1和2.2家族。

Establishment of cDNA of ductus arteriosus isolated from pig :Ductal tissue, aorta, pulmonary artery was isolated from 140 newborn pigs immediately after birth. Total RNA and then poly A RNA was isolated from the ductus, aorta, and pulmonary artery. cDNA was made from the RNA using RT-PCR. The cDNA was inserted into vector and cDNA library was established. The length of the inserted DNA was between 1.7 to 2.7 kb.Cloning Kv using the cDNA library :The cDNA library of pig ductus arteriosus was screened using Kv probes obtained from rat, mouse, and human beings. Kv 1, Kv 2, and Kv9.3 were cloned from the ductus arteriosus.Screening of the cDNA library using homology-based PCR :Primer was made using conserved part of Kv genes and PCR was performed using cDNA as a template. PCR products were compared between the ductus, aorta, and pulmonary artery. PCR products amplified only in the ductus was chosen and cloned. Kv families expressed in the ductus were cloned and they belong to 1.5, 2.1 and 2.2 family.