中文摘要：

应用PCR和RACE技术,以细菌脂多糖（LPS）刺激的草鱼头肾白细胞cDNA为模板,获得草鱼iNOS（In-ducible nitric oxide synthase）全长cDNA序列。该序列共4286 bp,编码含1080个氨基酸残基的蛋白。氨基酸序列比对分析发现草鱼iNOS与金鱼iNOS a和b、斑马鱼iNOS 2b以及鲤鱼iNOS高度保守,序列一致性均大于80%;它们的血红素、四氢生物蝶呤、钙调蛋白、FMN、FAD和NADH等辅因子结合区域也十分保守。用NJ法对新获得的序列和其它脊椎动物NOS的编码序列进行进化分析证实它属于诱导型NOS。在此基础上,运用基因步移技术分离草鱼iNOS基因的启动子序列,共有1978 bp。生物信息学分析发现该序列含有包括GR、AP1、C/EBP、ER、YY1、IRF1和IL-6 REBP在内的多个转录因子的潜在结合位点。

英文摘要：

The full length cDNA of grass carp iNOS was isolated from LPS-stimulated grass carp head kidney leucocytes by using PCR amplification coupling to 5＇-and 3＇-RACE.This cDNA was 4286 bp in length which encodes a protein with 1080 amino acid residues.Amino acid alignment showed that the identities between grass carp iNOS and goldfish iNOS,zebrafish iNOS2b and common carp iNOS were all more than 80%.The cofactor-binding sites for heme,tetrahydrobiopterin,calmodulin,FMN,FAD pyrophosphate,FAD isoalloxazine,NADPH ribose,NADPH adenine and NADPH are also highly conserved in these sequences.Phylogenetic analysis confirmed that this newly cloned iNOS is a member of inducible NOS.The promoter sequence of grass carp iNOS with 1978 bp was also obtained by using the Genome Walker Universal kit.Multiple transcription factors including GR,AP1,C/EBP,ER,YY1,IRF1 and IL-6 REBP binding sites were revealed by using transcription element search system in this study.