中国对虾3种Ⅱ型CHH家族神经肽基因的克隆及序列分析  

Cloning and Analysis of Three Genes Encoding Type Ⅱ CHH Family Neuropeptides from Fennropenaeus chinensis

作　　者：王在照[1] 相建海[1] 

机构地区：[1]中国科学院海洋研究所实验海洋生物学重点实验室,青岛266071

出　　处：《Acta Genetica Sinica》2003年第10期961-966,共6页

基　　金：国家自然科学基金资助项目 (编号 :40 10 60 18);国家高技术研究发展计划 (863 )资助项目 (编号 :2 0 0 1AA62 112 0 );国家重点基础研究发展规划资助项目 (编号 :G19990 12 0 0 7)~~

摘　　要：根据中国对虾蜕皮抑制激素的全长cDNA序列设计引物 ,以中国对虾基因组DNA为模板 ,采用PCR等技术分别得到 3个扩增产物 :NP1、NP2和NP3,长度分别是 5 4 0bp、6 0 1bp和 82 6bp。NP1和NP2均由 2个外显子和 1个内含子组成 ,NP3由 3个外显子和 2个内含子组成。序列分析表明 ,NP1、NP2和NP3是中国对虾Ⅱ型CHH家族神经肽的基因片段。NP1、NP2和NP3的外显子分别与斑节对虾的神经肽Pem SGP C2、Pem SGP C1及中国对虾FenchMIH的mRNA相似性最高 ,同源性分别为 91 5 %、92 8%、88 9%。由此可以推测NP3是中国对虾MIH的基因序列 。On the basis of sequence similarity,the crustean hy perglycemic hormone (CHH) family peptides have been classifed into two types of hormones:type Ⅰand type Ⅱ.Molt-inhibiting hormone (MIH) is a neuropeptide mem ber of type Ⅱ CHH family.Molting in shrimp is controlled by MIH and ecdysone.By inhibiting the synthesis of ecdysone in the Y-organ,MIH indirectly suppresses the molting activity of shrimp.In this study,we reported the cloning and charact er ization of 3 gene fragments encoding type Ⅱ CHH family neuropeptides of the shr imp Fennropenaeus chinensis.According to the complementary DNA sequence of t he molt-inhibiting hormone of Fennropenaeus chinensis,3 primers were design ed and synthesized.MP1 and MP2 are sense primers,and MP3 is anti-sense primer.P olymerase chain reaction was performed using genomic DNA of Fennropenaeus chin ensis as template.Three PCR products were obtained using pimers MP1 and MP3.Th eir sizes are about 600 bp,850 bp,1050 bp,respectively.A 580 bp PCR product was obta ined using primers MP2 and MP3.All the 4 PCR products were cloned into pMD18-T vector.The recombinant clones were sequenced using ABI 310 Genetic Analyzer.Afte r sequencing,all the DNA sequences were searched in the GenBank by Blast program to find similar gene sequences.The searching results revealed 3 DNA fragment se quences were of high similarity with CHH family neuropeptide genes from various crustean species.The 3 DNA fragments were named as NP1,NP2,and NP3.Their sizes w ere 540 bp,601 bp,and 826 bp,respectively.Using the mRNA sequences with the most similarity to the 3 sequence fragments as reference,the gene structure of the 3 DNA fragment sequences was analyzed.The exons of 3 sequence fragments were alig ned with their similar sequences by Clustal W program.Both NP1 and NP2 consisted of 1 intron and 2 exons.NP3 consisted of 2 introns and 3 exons.Sequence analys is suggested that these 3 products belonged to sequence fragments of neuropeptid e gene of type Ⅱ crustacean hyperglycemic hormone family.The exons

关 键 词：中国对虾 Ⅱ型CHH家族神经肽 

分 类 号：Q785[生物学—分子生物学]