title: Molecular cloning of hemocyanin cDNA from Fenneropenaeus chinensis and antimicrobial analysis of two C-terminal fragments.
abstract: Peptides derived from shrimp hemocyanin have antimicrobial properties. This is the first report of hemocyanin cDNA (FCHc) cloned from Fenneropenaeus chinensis and recombinant expression of two C-terminal fragments. Based on sequence analysis of Fenneropenaeus chinensis hemocyanin FCHc, we subcloned two FCHc fragments by designing special primers. Two antimicrobial peptides (AMPs) were derived from FCHc (FCHc-C1 and FCHc-C2). The recombinant sequence of FCHc-C1 consisted of 207 bp encoding 69 amino acids and the recombinant sequence of FCHc-C2 consisted of 120 bp encoding 40 amino acids. The results of Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting indicated that recombinant FCHc-C1 and FCHc-C2 peptides (rFCHc-C1 and rFCHc-C2) were expressed successfully. An inhibition assay showed that FCHc-C1 and FCHc-C2 were anionic AMPs with antifungal and antibacterial activities.
authors.affiliation: R&D Center of Marine Biotechnology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao, 266071, People's Republic of China.
authors.forename: Chuwen
authors.initials: C
authors.lastname: Qiu
authors.forename: Jie
authors.initials: J
authors.lastname: Sun
authors.forename: Mei
authors.initials: M
authors.lastname: Liu
authors.forename: Baojie
authors.initials: B
authors.lastname: Wang
authors.forename: Keyong
authors.initials: K
authors.lastname: Jiang
authors.forename: Shujuan
authors.initials: S
authors.lastname: Sun
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Meng
authors.forename: Zuoyong
authors.initials: Z
authors.lastname: Luo
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics* / Q000494:pharmacology; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors / Q000235:genetics; D005998:Glycine / Q000031:analogs & derivatives; D006433:Hemocyanins / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000235:genetics*; D010843:Pichia / Q000187:drug effects; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D023181:Antimicrobial Cationic Peptides; D017931:DNA Primers; D018076:DNA, Complementary; D011994:Recombinant Proteins; D006433:Hemocyanins; D005998:Glycine; C100184:tricine