title: Molecular cloning and characterization of lactate dehydrogenase gene from Eimeria tenella.
abstract: Lactate dehydrogenase (LDH) is a key enzyme in the glycolytic pathway and is crucial for parasite survival. In this study, we cloned and expressed the LDH of Eimeria tenella (EtLDH). Real-time polymerase chain reaction and Western blot analysis revealed that the expression of EtLDH was developmentally regulated at the messenger RNA (mRNA) and protein levels. EtLDH mRNA levels were higher in second-generation merozoites than in other developmental stages (unsporulated oocysts, sporulated oocysts, and sporozoites). EtLDH protein expression levels were most prominent in second-generation merozoites, moderately expressed in unsporulated oocysts and sporulated oocysts, and weakly detected in sporozoites. Immunostaining with anti-recombinant EtLDH (rEtLDH) antibody indicated that EtLDH was mainly located in the anterior region in free sporozoites and became concentrated in the anterior region of intracellular sporozoites except for the apex after invasion into DF-1 cells. Specific staining of EtLDH protein was more intense in trophozoites and immature first-generation schizonts, but decreased in mature first-generation schizonts. Inhibition of EtLDH function using specific antibodies cannot efficiently reduce the ability of E. tenella sporozoites to invade host cells. These results suggest that EtLDH may be involved in glycolysis during the first-generation merogony stage in E. tenella and has little role in host invasion.
authors.affiliation: Shanghai Veterinary Research Institute, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Chinese Academy of Agricultural Science, No. 518 Ziyue Road, Minhang District, 200241, Shanghai, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Dong
authors.forename: Yange
authors.initials: Y
authors.lastname: Wang
authors.forename: Qiping
authors.initials: Q
authors.lastname: Zhao
authors.forename: Hongyu
authors.initials: H
authors.lastname: Han
authors.forename: Shunhai
authors.initials: S
authors.lastname: Zhu
authors.forename: Liujia
authors.initials: L
authors.lastname: Li
authors.forename: Youling
authors.initials: Y
authors.lastname: Wu
authors.forename: Bing
authors.initials: B
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D016786:Eimeria tenella / Q000201:enzymology* / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D007770:L-Lactate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D052940:Merozoites / Q000201:enzymology; D008969:Molecular Sequence Data; D033661:Oocysts / Q000201:enzymology; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D034101:Sporozoites / Q000201:enzymology
chemical_list: D015800:Protozoan Proteins; D011994:Recombinant Proteins; D007770:L-Lactate Dehydrogenase