cDNA cloning and expression of a collectin from red-spotted grouper (Epinephelus akaara)

Zhiwen Zhang; Shaoxiong Ding; Ying Wang; Yong Mao; Yongquan Su; Jun Wang

doi:10.1007/s00343-009-9291-z

Your Location：

Home >

Browse articles >

cDNA cloning and expression of a collectin from red-spotted grouper (Epinephelus akaara)

Updated：2023-05-16

- cDNA cloning and expression of a collectin from red-spotted grouper (Epinephelus akaara)
- Journal of Oceanology and Limnology Vol. 27, Issue 3, Pages: 543-549(2009)
- Affiliations：
  
  College of Oceanography and Environmental Science Xiamen University
- Author bio：
- Funds：
- DOI：10.1007/s00343-009-9291-z
  CLC：
- Online First：07 November 2009，
  
  Published：2009
- Accepted：
Scan QR Code
Zhang, Z., Ding, S., Wang, Y. et al. cDNA cloning and expression of a collectin from red-spotted grouper (Epinephelus akaara)., Chin. J. Ocean. Limnol. 27, 543–549 (2009). https://doi.org/10.1007/s00343-009-9291-z
DOI：

Zhang, Z., Ding, S., Wang, Y. et al. cDNA cloning and expression of a collectin from red-spotted grouper (Epinephelus akaara)., Chin. J. Ocean. Limnol. 27, 543–549 (2009). https://doi.org/10.1007/s00343-009-9291-z DOI：

摘要

Abstract

Lectins play a crucial role in the innate immunity of invertebrates and vertebrates by recognizing and disposing of pathogens. We obtained the complete cDNA of a C-type lectin (EALec1) from Epinephelus akaara using RACE. The complete EALec1 cDNA sequence was 827 bp. The 5-UTR and 3-UTR were 28 bp and 151 bp

respectively

in length. The sequence also contained a polyadenylation signal AATAAA and a poly(A) tail. The EALec1 cDNA encodes polypeptides with 215 amino acids

including a signal peptide of 31 amino acids. The protein has a cysteine-rich region at the N terminal

a collagenous region characterized by G-X-Y repeats

a neck region

and a typical carbohydrate-recognition domain (CRD)

indicating that EALec1 is a collectin. The key recognition positions of this CRD are EPD

isolated for the first time in fish. These are likely the interim types

between mannan-binding lectin and galactose-binding lectin. We evaluated the expression pattern of EALec1 in 12 different tissues using RT-PCR. EALec1 was expressed in all tissues

though at different levels. In addition

we inserted EALec1 into an expression vector (pET-28a) for transformation into the BL21 engineering bacteria. Based on enzyme digestion and sequencing of the positive clone

we successfully constructed the EALec1 recombinant expression vector.

关键词

Keywords

references

Dah M R, Thiel S, Matsushita M. 2001. MASP-3 and its association with distinct complexes of the mannan-binding lectin complement activation pathway. Immunity, 15: 127–135.

Ellis A E. 2001. Innate host defense mechanisms of fish against viruses and bacteria. Dev. Comp. Immunol., 25(8–9): 827–839.

Ewart K V, Johnson S C, Ross N W. 2001. Lectins of the innate immune system and their relevance to fish health. ICES Journal of Marine Science, 58: 380–385.

Hans-Jurgen H, Kenneth B M. 1994. Collectins — soluble proteins containing collagenous regions and lectin domains -and their roles in innate immunity. Protein Science, 3: 1 143–1 158.

Jackson A N, C A, McLure R L Dawkins, Keating P J. 2007. Mannose binding lectin (MBL) copy number polymerphism in Zebrafish (D. rerio) and identification of haplotypes resistant to L. anguillarum. Immunogenetics, 59(11): 861–872.

Kondo H, Yeu T A, Hirono I. 2007. Identification of a novel C-type lectin gene in Japanese flounder. Paralichthys olivaceus. Fish Shellfish Immunol., 23(5): 1 089–1 094.

Medzhitov R, Janeway J C A. 1998. Innate immune recognition and control of adaptive immune responses. Seminars in Immunology, 10: 351–353.

Nakao M, Kajiya T, Sato Y, Somamoto T. 2006. Lectin pathway of bony fish complement: identification of two homologs of the mannose-binding lectin associated with MASP2 in the common carp (Cyprinus carpio). Immunol., 177(8): 5 471–5 479.

Nikolakopoulou K, Zarkadis I K. 2006. Molecular cloning and characterization of two homologues of Mannose-Binding Lectin in rainbow trout. Fish Shellfish Immunol., 21(3): 305–314.

Panagos P G, Dobrinski K P, Chen X. 2006. Immune-related, lectin-like receptors are differentially expressed in the myeloid and lymphoid lineages of zebrafish. Immunogenetics, 58(1): 31–40.

Russell S, Lumsden J S. 2005. Function and heterogeneity of fish lectins. Vet. Immunol. Immunopathol., 108(1–2): 111–120.

Russell S, Young K M, Smith M. 2008. Cloning, binding properties, and tissue localization of rainbow trout (Oncorhynchus mykiss) ladderlectin. Fish Shellfish Immunol., 24(6): 669–683.

Sharon E, Sharon N, NirB T. 2003. Evolutionary analysis reveals collective properties and specificity in the C-Type lectin and lectin-like domain superfamily. PROTEINS: Structure, Function, and Bioinformatics, 53: 44–55.

Sharon N, Lis H. 1989. Lectins as cell recognition molecules. Science, 246: 227–234.

Tsutsui S, Tasumi S, Suetake H. 2006. Carbohydrate-binding site of a novel mannose-specific lectin from fugu (Takifugu rubripes) skin mucus. Comp. Biochem. Physiol. B. Biochem. Mol. Biol., 143(4): 514–519.

Tsutsui S, Tasumi S, Suetake H. 2005. Demonstration of the mucosal lectins in the epithelial cells of internal and external body surface tissues in pufferfish (Fugu rubripes). Dev. Comp. Immunol., 29(3): 243–253.

Vitved L, Holmskov U, Koch C. 2000. The homologue of mannose-binding lectin in the carp family Cyprinidae is expressed at high level in spleen, and the deduced primary structure predicts affinity for galactose. Immunogenetics, 51(11): 955–964.

Wang H, Song L, Li C, Zhao J. 2007. Cloning and characterization of a novel C-type lectin from Zhikong scallop Chlamys farreri. Mol. Immunol., 44(5): 722–731.

Wang N, Whang I, Lee J. 2008. A novel C-type lectin from abalone, Haliotis discus discus, agglutinates Vibrio alginolyticus. Developmental and Comparative Immunology, 32: 1 034–1 040.

Yuichi E, Momoe T, Teizo F. 2006. Lectin complement system and pattern recognition. Immunobiology, 211: 283–293.

Zelensky A N, Gready J E. 2005. The C-type lectin-like domain superfamily. FEBS J., 272(24): 6 179–6 217.

Zelensky A N, Gready J E. 2004. C-type lectin-like domains in Fugu rubripes. BMC Genomics, 5(1): 51.

Views

Downloads

CSCD

Alert me when the article has been cited

Submit

Tools

Publicity Resources

Cloning and expression of prion protein encoding gene of flounder (Paralichthys olivaceus)

The role of Smad6 in immunity of the pearl oyster Pinctada fucata martensii

Cloning of catalase gene and antioxidant genes in Scophthalmus maximus response to metalloprotease of Vibrio anguillarum stress

Triterpenoid saponin biosynthesis genes and their expression patterns during the development of sea cucumber Apostichopus japonicus

Isolation, cloning and sequencing of AFLP markers related to disease-resistance traits inFenneropenaeus chinensis

Related Author

Jindong Zan

Jinxing Zhang

Xiuqin Sun

Zhiwen Zhang

Maoxian HE

Hanzhi XU

Huiru LIU

Meng XU

Related Institution

The First Institute of Oceanography State Oceanic Administration of China

College of Marine Life Sciences Ocean University of China

Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou)

CAS Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Provincial Key Laboratory of Applied Marine Biology, South China Sea Institute of Oceanology, Chinese Academy of Sciences

Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences

Address：Editorial Office of JOL, 88 Haijun Rd. Qingdao, 266000, China
Tel：+86-532-82898754 Email：jol@qdio.ac.cn
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰