三疣梭子蟹蜕皮抑制激素cDNA的克隆与序列分析  

Molecular cloning and sequence analysis of cDNA encoding molt-inhibiting hormone from Portunus trituberculatus

作　　者：朱冬发[1] 沈建明[1] 杨济芬[1] 苏青[1] 

机构地区：[1]宁波大学应用海洋生物技术教育部重点实验室,浙江宁波315211

出　　处：《动物学报》2008年第6期1112-1118,共7页ACTA ZOOLOGICA SINICA

基　　金：国家自然科学基金(No.40576068);长江学者和创新团队发展计划(No.IRT0734);浙江省科技计划(No.2008C22048)资助~~

摘　　要：Molt-inhibiting hormone(MIH),which belongs to the crustacean hyperglycemic hormone(CHH)neuropeptide family,inhibits the synthesis of ecdysteroids by Y-organs.The CHH family is divided into two major groups,designated Ⅰ and Ⅱ.Group Ⅰ is composed of all CHH peptides,and Group Ⅱ is composed of most MIHs,gonad-inhibiting hormone(GIH)and mandibular organ-inhibiting hormone(MOIH).The Group Ⅰ peptides consist of a signal sequence,a CHH precursor related peptide(CPRP)and a mature hormone.The Group Ⅱ peptides have only a signal sequence and a mature hormone.The full-length molt-inhibiting hormone cDNA of Portunus trituberculatus(Pot-MIH)was cloned by the reverse transcription-polymerase chain reaction(RT-PCR)and the rapid amplification of cDNA ends(RACE).The degenerate nucleotide primers was designed based on the amino acid sequence of other crustacean MIHs.The full-length sequence of Pot-MIH cDNA(GenBank accession number:EU284117)is 1572 bp in size and consists of a 210 bp 5’-untranslated region(5’-UTR),a 342 bp open reading frame and a 3’-untranslated region(3’-UTR).The deduced polypeptide consisted of a 78-amino acid mature MIH peptide and a 35-amino acid signal ptptide.The mature Pot-MIH peptide shared twelve conserved residues(Cys7,Gly12,Arg14,Cys24,Asp26,Cys27,Asn29,Arg32,Cys40,Cys44,Phe50,and Cys53)with other crustacean MIHs.The mature Pot-MIH peptide had the Gly12 residue and the precursor lacked a CPRP sequence,both of which are characteristic of all Group Ⅱ peptides.Alignment of the amino acid sequence deduced from the full-length Pot-MIH cDNA with MIH amino acid sequences of other crustaceans revealed that they had very high identity.The identities between the putative Pot-MIH amino acid sequence with that of MIH of P.pelagicus,Callinectes sapidus,Charybdis feriatus,Cancer pagurus,C.magister,Carcinus maenas and Gecarcinus lateralis were 97%,95%,90%,82%,81%,77% and 60% respectively.Phylogenetic analysis indicates that the putative Pot-MIH cluster with other crustacean MIHs and GIHs.Molt-inhibiting hormone （MIH）, which belongs to the crustacean hyperglycemic hormone （CHH） neuropeptide family, inhibits the synthesis of ecdysteroids by Y-organs. The CHH family is divided into two major groups, designated Ⅰ and Ⅱ. Group is composed of all CHH peptides, and Group Ⅱ is composed of most MIHs, gonad-inhibiting hormone （GIH） and mandibular organ-inhibiting hormone （MOIH）. The Group Ⅰ peptides consist of a signal sequence, a CHH precursor related peptide （CPRP） and a mature hormone. The Group Ⅱ peptides have only a signal sequence and a mature hormone. The full-length molt-inhibiting hormone cDNA of Portunus trituberculatus （Pot-MIH） was cloned by the reverse transcription-polymerase chain reaction （RTPCR） and the rapid amplification of cDNA ends （RACE）. The degenerate nucleotide primers was designed based on the amino acid sequence of other crustacean MIHs. The full-length sequence of Pot-MIH cDNA （GenBank accession number： EU284117） is 1572 bp in size and consists of a 210 bp 5′-untranslated region （5′-UTR）, a 342 bp open reading frame and a 3′- untranslated region （3′-UTR）. The deduced polypeptide consisted of a 78-amino acid mature MIH peptide and a 35-amino acid signal ptptide. The mature Pot-MIH peptide shared twelve conserved residues （Cys^7 , Gly^12 , Arg^34 , Cys^24 , Asp^26 , Cys^27 , Asn^29 , Arg^32 , Cys^40 , Cys^44, Phe^50, and Cys53） with other crustacean MIHs. The mature Pot-MIH peptide had the Gly^12 residue and the precursor lacked a CPRP sequence, both of which are characteristic of all Group Ⅱ peptides. Aligmnent of the amino acid sequence deduced from the full-length Pot-MIH cDNA with MIH amino acid sequences of other crustaceans revealed that they had very high identity. The identities between the putative Pot-MIH amino acid sequence with that of MIH of P. pelagicus, Callinectes sapidus, Charybdis feriatus, Cancer pagurus, C. magister, Carcinus maenas and Gecarcinus lateralis were 97 % , 95 % , 90 % , 82 % , 8

关 键 词：三疣梭子蟹 蜕皮抑制激素 CDNA克隆 序列分析 

分 类 号：Q78[生物学—分子生物学]