title: Evidences of abundant hemocyanin variants in shrimp Litopenaeus vannamei.
abstract: Hemocyanin (HMC) is a multifunctional immune molecule present in mollusks and arthropods and functions as an important antigen non-specific immune protein. Our previous evidences demonstrated that Litopenaeus vannamei HMC might display extensive molecular diversities. In this study, bioinformatics analysis showed dozens of variant sequences of the HMC subunit with higher molecular weight from L. vannamei (LvHMC). Three variant fragments, named as LvHMCV1-3, which shared 85-99% nucleotide identity with that of the classical form of LvHMC (AJ250830.1), were cloned and characterized. Spatial expression profiles showed that LvHMCV1-3 had different tissue-specific distribution, which were affected by stimulation with six pathogenic bacteria, including Escherichia coli K12, Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio fluvialis, Streptococcus pyogenes and Staphylococcus aureus, with each variant fragment showing a specific stress pattern to different bacterial pathogens. Full length cDNA of LvHMCV3 was further cloned and characterized. The deduced amino acid sequence shared 92% identity with that of LvHMC, possessed a conserved structure characteristic of the HMC family and could be clustered into one branch along with other arthropod HMC in a phylogenetic tree. In addition, the recombinant protein of LvHMCV3 (rLvHMCV3) showed obvious agglutination activities against three aquaculture pathogenic bacteria including E. coli K12, V. parahaemolyticus and S.aureus at concentrations ranging from 31.25-62.5g/mL. It also showed obvious antibacterial activity against V. parahaemolyticus at concentrations 0.02-0.5mg/mL, and possessed the best inhibitive effects compared with those of rLvHMCV4 and rLvHMC. Co-injection of V. parahaemolyticus and rLvHMCV3 in L. vannamei showed significant decrease of the mortality rate at 24-72h after injection. Therefore, these studies suggested that L. vannamei had abundant HMC variants, which possessed obvious resistance to pathogenic infection and might specifically target on different pathogens in shrimp.
authors: Department of Biology and Guangdong Provincial Key laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China.
authors: Xianliang
authors: X
authors: Zhao
authors: Department of Biology and Guangdong Provincial Key laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China.
authors: Lingling
authors: L
authors: Guo
authors: Department of Biology and Guangdong Provincial Key laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China.
authors: Xin
authors: X
authors: Lu
authors: Department of Biology and Guangdong Provincial Key laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China.
authors: Hui
authors: H
authors: Lu
authors: Department of Biology and Guangdong Provincial Key laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China.
authors: Fan
authors: F
authors: Wang
authors: Department of Biology and Guangdong Provincial Key laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China.
authors: Mingqi
authors: M
authors: Zhong
authors: Department of Biology and Guangdong Provincial Key laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China.
authors: Jiehui
authors: J
authors: Chen
authors: Department of Biology and Guangdong Provincial Key laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China. Electronic address: zhangyl@stu.edu.cn.
authors: Yueling
authors: Y
authors: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019295:Computational Biology; D014644:Genetic Variation / Q000276:immunology; D006433:Hemocyanins / Q000235:genetics / Q000276:immunology*; D033561:Penaeidae / Q000235:genetics / Q000276:immunology*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment
chemical_list: D006433:Hemocyanins
keywords: Hemocyanin; Litopenaeus vannamei; Pathogen; Resistance; Specificity; Variant