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Molecular characterization and expression analysis of chitinase (Fcchi-3) from Chinese shrimp, Fenneropenaeus chinensis
Zhang, Jiquan1; Sun, Yuying2; Li, Fuhua1; Huang, Bingxin1; Xiang, Jianhai1
2010-04-01
Source PublicationMOLECULAR BIOLOGY REPORTS
Volume37Issue:4Pages:1913-1921
SubtypeArticle
AbstractComplementary DNA (cDNA) and genomic DNA, including flanking regions of the chitinase gene (Fcchi-3) of Fenneropenaeus chinensis, were cloned and sequenced. Fcchi-3 was found to have 92.0 and 91.4% identity at the cDNA level to that of Litopenaeus vannamei and Marsupenaeus japonicus, respectively. The predicted amino acid sequence had an overall similarity with a comparable region of L. vannamei (96.8%) and M. japonicus (93.4%). Based on the cDNA sequence, the genomic structure of the gene was characterized. Sequence analysis revealed that the Fcchi-3 gene was composed of seven exons with 411, 252, 186, 132, 171, 117 and 135 bp and six introns with 232, 196, 121, 90, 159 and 157 bp. Analysis by RT-PCR revealed that Fcchi-3 was a hepatopancreas specific gene. Semi-quantitative RT-PCR analysis revealed that Fcchi-3 transcript was down-regulated significantly in response to the challenge of WSSV at 5 h post-injection and then came back to normal level at 37 h. A fusion protein containing Fcchi-3 was produced and the purified recombinant protein exhibited similar biological function. The result of identification through LC-ESI-MS showed that three peptide fragments (-MAADPVLR-, -ATIDPAYNVPELSK- and -AILAVGGWNEGSPK-) of the recombinant protein were identical to the corresponding sequence of L. vannamei chitinase-3. The recombinant Fcchi-3 could degrade the colloid chitin confirming that the recombinant protein is actually the chitinase.
KeywordFenneropenaeus Chinensis Chitinase Recombinant Expression Lc-esi-ms
DOI10.1007/s11033-009-9633-0
Indexed BySCI
Language英语
WOS IDWOS:000275161700029
Citation statistics
Cited Times:20[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/32810
Collection实验海洋生物学重点实验室
Affiliation1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China
2.Huaihai Inst Technol, Jiangsu Key Lab Marine Biotechnol, Lianyungang 222005, Peoples R China
First Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Recommended Citation
GB/T 7714
Zhang, Jiquan,Sun, Yuying,Li, Fuhua,et al. Molecular characterization and expression analysis of chitinase (Fcchi-3) from Chinese shrimp, Fenneropenaeus chinensis[J]. MOLECULAR BIOLOGY REPORTS,2010,37(4):1913-1921.
APA Zhang, Jiquan,Sun, Yuying,Li, Fuhua,Huang, Bingxin,&Xiang, Jianhai.(2010).Molecular characterization and expression analysis of chitinase (Fcchi-3) from Chinese shrimp, Fenneropenaeus chinensis.MOLECULAR BIOLOGY REPORTS,37(4),1913-1921.
MLA Zhang, Jiquan,et al."Molecular characterization and expression analysis of chitinase (Fcchi-3) from Chinese shrimp, Fenneropenaeus chinensis".MOLECULAR BIOLOGY REPORTS 37.4(2010):1913-1921.
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