Total RNA was extracted from eyestalks of shrimp Penaeus chinensis . Eyestalk cDNA was obtained from total RNA by reverse transcription. Reverse transcriptase polymerase chain reaction (RT PCR) was initiated using eye...Total RNA was extracted from eyestalks of shrimp Penaeus chinensis . Eyestalk cDNA was obtained from total RNA by reverse transcription. Reverse transcriptase polymerase chain reaction (RT PCR) was initiated using eyestalk cDNA and degenerate primers designed from the amino acid sequence of molt inhibiting hormone from shrimp Penaeus japonicu s. A specific cDNA was obtained and cloned into a T vector for sequencing. The cDNA consisted of 201 base pairs and encoding for a peptide of 67 amino acid residues. The peptide of P. chinensis had the highest identity with molt inhibiting hormones of P. japonicus . The cDNA could be a partial gene of molt inhibiting hormones from P. chinensis . This paper reports for the first time cDNA encoding for neuropeptide of P. chinensis .展开更多
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 ...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.展开更多
文摘Total RNA was extracted from eyestalks of shrimp Penaeus chinensis . Eyestalk cDNA was obtained from total RNA by reverse transcription. Reverse transcriptase polymerase chain reaction (RT PCR) was initiated using eyestalk cDNA and degenerate primers designed from the amino acid sequence of molt inhibiting hormone from shrimp Penaeus japonicu s. A specific cDNA was obtained and cloned into a T vector for sequencing. The cDNA consisted of 201 base pairs and encoding for a peptide of 67 amino acid residues. The peptide of P. chinensis had the highest identity with molt inhibiting hormones of P. japonicus . The cDNA could be a partial gene of molt inhibiting hormones from P. chinensis . This paper reports for the first time cDNA encoding for neuropeptide of P. chinensis .
文摘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.
