Molecular cloning and characterization of lactate dehydrogenase gene from Eimeria tenella
| Autor: | Qiping Zhao, Liujia Li, Bing Huang, Hongyu Han, Youling Wu, Shunhai Zhu, Hui Dong, Yange Wang |
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| Rok vydání: | 2014 |
| Předmět: |
Molecular Sequence Data
Protozoan Proteins Biology Molecular cloning Real-Time Polymerase Chain Reaction Eimeria chemistry.chemical_compound Western blot Lactate dehydrogenase parasitic diseases medicine Animals Amino Acid Sequence Cloning Molecular Regulation of gene expression Messenger RNA L-Lactate Dehydrogenase General Veterinary medicine.diagnostic_test Merozoites Oocysts Gene Expression Regulation Developmental General Medicine biology.organism_classification Molecular biology Recombinant Proteins Infectious Diseases Real-time polymerase chain reaction chemistry Sporozoites Insect Science Parasitology Eimeria tenella Immunostaining |
| Zdroj: | Parasitology Research. 113:2915-2923 |
| ISSN: | 1432-1955 0932-0113 |
| DOI: | 10.1007/s00436-014-3953-2 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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