[Expression and cloning of two novel genes induced by ischemia and reperfusion].
OBJECTIVE
To identify and clone the differentially expressed genes in brief ischemia and reperfusion myocardium.


METHODS
Ischemia and reperfusion were induced by repeated brief ligation of the porcine left anterior descending coronary artery. Total RNA which was isolated from myocardium subjected ischemia and reperfusion was used for mRNA differential display. After cloning and sequencing the cDNA fragments which showed change in expression, their expression were further confirmed by Northern-Blot analysis.


RESULTS
Two differentially expressed cDNAs (W12 and W28) were identified and cloned. Their expression were subsequently confirmed to be truly differentially expressed. The expression of both genes in ischemia and reperfusion myocardium was obvious higher than that in nonischemia and reperfusion: W12 expression level was 2-fold (P < 0.05), and W28 expression level 1.9-fold (P < 0.05). In addition, mRNAs of W12 and W28 were existed in all tested organs including heart, liver, lung, kidney, spleen, intestine, brain and skeletal muscle. DNA sequencing analysis showed that there was no homology between W12, W28 and known genes, implying that they would represent novel gene respectively.


CONCLUSIONS
Two novel genes induced by ischemia and reperfusion are identified, cloned and confirmed.
Wang
G P
GP
Central Laboratory, Xiangya Hospital, Central South University, Changsha 41008, China.
Cheng
B L
BL
Cai
W J
WJ
D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D015428:Myocardial Reperfusion Injury / Q000235:genetics*; D009206:Myocardium / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011897:Random Allocation; D013556:Swine, Miniature
D018076:DNA, Complementary; D012333:RNA, Messenger