Expression and characterization of the TsNas36 protein of Trichinella spiralis
10.16303/j.cnki.1005-4545.2025.06.16
- VernacularTitle:旋毛虫TsNas36蛋白的表达和生物学特性分析
- Author:
Guangquan SI
1
;
Junpeng SONG
;
Qingbo LYU
;
Xue BAI
;
Yang WANG
;
Xiaolei LIU
;
Lixi-ao ZHANG
Author Information
1. 吉林大学动物医学学院人畜共患传染病重症诊治全国重点实验室/人兽共患病研究教育部重点实验室/人兽共患病研究所,吉林长春 130062
- Publication Type:Journal Article
- Keywords:
Trichinella spiralis;
nematode astacins;
Zinc metalloproteinase;
TsNas36;
prokaryotic expression;
immunofluorescence
- From:
Chinese Journal of Veterinary Science
2025;45(6):1225-1232,1242
- CountryChina
- Language:Chinese
-
Abstract:
Trichinella spiralis zinc metalloproteinase NAS-36 gene(TsNas36)is a member of the zinc metalloproteinase family found in excretory secretory products(ESP)of T.spiralis.In this study,TsNas36 gene was cloned and expressed,and its biological characteristics and temporal and spatial characteristics were identified.These results provide a theoretical and material basis for ex-ploring the biological function of TsNas36 gene.Bioinformatics analysis showed that TsNas36 was 470 amino acids(AA)in length with a molecular weight of about 54.69 kDa,no transmembrane region,and contained a signal peptide(1-20 AA),an Astacins domain(116-320 AA)and a CUB domain(355-470 AA).There were five active site residues located at amino acids 216(His),217(Glu),220(His),226(His)and 275(Tyr).The expression plasmid pET-28a(+)/TsNas36 was constructed and induced to express in E.coli BL21(DE3)to obtain the recombinant protein rTs-Nas36.The recombinant protein was used to immunize rabbits to obtain anti-rTsNas36 polyclonal antibody serum.Indirect ELISA results showed that the antibody titer reached 1∶105.qRT-PCR and Western blot results showed that the transcription levels of TsNas36 were significantly higher in newborn larvae(NBL)than in adult worm(AW)and muscle larva(ML)stages.Immunofluo-rescence results showed that TsNas36 was only localized in the epidermis of NBL.In summary,this study characterized the biological characteristics of the TsNas36 gene and found that this gene is highly period-specific and may be involved in the unique developmental process of NBL.
