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| 凡纳滨对虾细胞色素P450家族新基因CYP4V18的克隆及KK-42对其转录的抑制 | |
| 引用本文： | 夏西超,王文锋,杨洪,宁黔冀.凡纳滨对虾细胞色素P450家族新基因CYP4V18的克隆及KK-42对其转录的抑制[J].中国水产科学,2012,19(5):767-774. |
| 作者姓名： | 夏西超 王文锋 杨洪 宁黔冀 |
| 作者单位： | 1. 河南师范大学生命科学学院,河南新乡453007 2. 南阳医学高等专科学校基础医学部,河南南阳473061 3. 新乡医学院,河南新乡453003 |
| 基金项目： | 国家自然科学基金项目(30940008);高等学校博士学科点专项科研基金(20094104110003);河南省高校科技创新人才支持计划(2009HASTIT022) |
| 摘 要： | 为探讨咪唑类物质KK-42促进凡纳滨对虾(Litopenaeaus vannamei)生长的分子机制,本研究克隆出与蜕皮和生长相关的CYP4V18基因的全序列,研究了该基因的时空表达,并结合测定血淋巴20-羟基蜕皮甾酮(20E)滴度。将体长3.5～5.0 cm凡纳滨对虾幼虾随机分为2组,分别用1.95×10-4mol/L的KK-42溶液或不含KK-42的溶液浸泡处理1 min,之后在相同条件下常规饲养。CYP4V18 mRNA水平定量分析采用real-time PCR,血淋巴中20E滴度测定采用反相高效液相色谱法。结果显示,CYP4V18开放阅读框为1 545 bp,编码515个氨基酸,包含CYP450保守序列;系统进化显示,CYP4V18与CYP4家族基因的亲缘关系最近。CYP4V18 mRNA水平在肝胰腺最高,分别是脑、Y-器官和肌肉的4、26和32倍。KK-42处理可显著抑制CYP4V18表达,在第2、4和6天分别降低了72.3%(P<0.05)、82.6%(P<0.05)和187.7%(P<0.01)。与同期对照组相比,KK-42处理之后血淋巴20E滴度在第2、4和6天分别升高1.7%、6.5%和13.5%。结果表明,KK-42可显著抑制凡纳滨对虾肝胰腺CYP4V18转录,升高血淋巴20E滴度,这可能是KK-42促生长机制之一。 |
| 关 键 词： | 凡纳滨对虾 CYP4V18基因 KK-42 表达下调 |
| 修稿时间： | 2012/10/15 0:00:00 |
A novel CYP450 gene (CYP4V18) of the shrimp Litopenaeaus vannamei: Cloning and down-regulation of CYP4V18 expression by the imidazole derivative KK-42 |
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| XIA Xichao,WANG Wenfeng,YANG Hong,NING Qianji.A novel CYP450 gene (CYP4V18) of the shrimp Litopenaeaus vannamei: Cloning and down-regulation of CYP4V18 expression by the imidazole derivative KK-42[J].Journal of Fishery Sciences of China,2012,19(5):767-774. | |
| Authors: | XIA Xichao WANG Wenfeng YANG Hong NING Qianji |
| Institution: | 1.College of Life Sciences,Henan Normal University,Xinxiang 453007,China; 2.Nanyang Medical College Nanyang 473061,China; 3.Xinxiang Medical University,Xinxiang 453002,China |
| Abstract: | 20E) plays a role in the direct regulation of crustacean molting and growth. The synthesis and degradation of 20E is catalyzed by cytochrome P450 enzymes (CYP enzymes). We attempted to determine the molecular mechanism for the growth promoting effect of KK-42 in) (body length 3.55.0 cm) were exposed to a solution containing KK-42 (1.95×10 mol/L) or to a control solution without KK-42 then cultured normally. A single was cloned and its spatiotemporal expression was analyzed using real-time PCR. The hemolymph 20E titer was measured by reverse-phase high performance liquid chromatography. The full-length cDNA contained a 1 545 bp open reading frame encoding a predicted protein of 515 amino acids which shares common characteristics with the cytochrome P450 superfamily. The expression of mRNA was highest in the hepatopancreas. Following KK-42 treatment, expression of hepatopancreas decreased significantly to >72.3 % that of the control group and was that of the control on day 6. Administration of KK-42 resulted in an increase in hemolymph 20E titers compared to that of control animals. Our results suggest that the inhibition of hepatopancreas expression and the increase in hemolymph 20E titer following KK-42 administration may explain the growth promoting effect of KK-42 in |
| Keywords: | Litopenaeaus vannamei CYP4V18 gene KK-42 down-regulation |
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