Park C S, Pardhasaradhi K, Gianotti C, Villegas E, Krishna G
Section of Chemical Pharmacology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892-1760.
Biochem Biophys Res Commun. 1994 Nov 30;205(1):85-91. doi: 10.1006/bbrc.1994.2633.
The present study provided evidence for the presence of two forms of nitric oxide synthase(NOS) gene in the human retina. Expression of retinal constitutive type(rbNOS) and inducible type(riNOS) of NOS was detected in human retinal poly A+RNA by reverse transcriptase polymerase chain reaction (RT-PCR) method. The deduced amino acid sequence of the human retinal rbNOS showed more than 99% homology with human brain bNOS and that of riNOS was identical to the chondrocytes inducible iNOS with the exception for one amino acid. These differences in amino acid sequences of rbNOS and riNOS, with their counterparts in human brain and human chondrocytes sequences, were only in the non-cofactor binding sites. Northern blot analysis of the human retinal poly A+RNA and total RNA, using the PCR-amplified riNOS probe revealed the existence of riNOS message with the appearance of the band with the expected size of 4.4kb, while the message for rbNOS was not detectable. This was the first report of the deduced nucleotide sequence identification of two NOS genes from a human tissue, while there had been earlier reports from culture cells.
本研究为人类视网膜中存在两种形式的一氧化氮合酶(NOS)基因提供了证据。通过逆转录聚合酶链反应(RT-PCR)方法,在人视网膜多聚腺苷酸(poly A+)RNA中检测到了视网膜组成型(rbNOS)和诱导型(riNOS)NOS的表达。人视网膜rbNOS推导的氨基酸序列与人类脑bNOS显示出超过99%的同源性,而riNOS的推导氨基酸序列除一个氨基酸外与软骨细胞诱导型iNOS相同。rbNOS和riNOS氨基酸序列与其在人类脑和人类软骨细胞序列中的对应序列的这些差异仅存在于非辅因子结合位点。使用PCR扩增的riNOS探针,对人视网膜多聚腺苷酸(poly A+)RNA和总RNA进行Northern印迹分析,结果显示存在预期大小为4.4kb的条带,表明存在riNOS信息,而未检测到rbNOS信息。这是首次从人类组织中推导鉴定出两种NOS基因的核苷酸序列的报告,而此前已有来自培养细胞的相关报告。