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

猪分离动脉导管cDNA的建立：从140头刚出生的猪身上分离导管组织、主动脉、肺动脉。从导管、主动脉和肺动脉中分别分离总RNA和多聚RNA。利用RT-PCR从RNA中提取cDNA。将cDNA插入载体，建立cDNA文库。插入的DNA长度在1.7 ~ 2.7 kb之间。利用cDNA文库克隆Kv：利用大鼠、小鼠和人的Kv探针筛选猪动脉导管cDNA文库。kv1、kv2和Kv9.3克隆自动脉导管。同源PCR筛选cDNA文库：以Kv基因保守部分为引物，以cDNA为模板进行PCR。对导管、主动脉和肺动脉的PCR产物进行比较。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，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.