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31.
Rongrong ZHANG Ting LEI Yanmei QI Peng LEI Zhilong CHEN Xiaodong CHEN Zaiqing YANG 《Animal Science Journal》2010,81(4):425-431
Adenosine diphosphate‐ribosylation factors (Arfs) are a family of guanosine triphosphate‐binding proteins involved in fundamental biological processes including secretion, endocytosis, phagocytosis, cytokinesis, cell adhesion and tumor cell invasion. We report here the molecular cloning, chromosome localization and expression analysis of porcine Arf1–6, of which Arf1–3 (Class I) have >93% similarity to each other and encode nearly similar proteins with 181 amino acids in length. Arf4 and Arf5 (Class II) are 78–81% homologous to Class I Arfs and both encode a protein of 180 amino acids, Arf6 (Class III) shows 64–68% homology to the other Arfs and encodes 175 amino acids. With radiation hybrid mapping, porcine Arf1–6 are assigned to chromosomes 14q21‐q22, 12p14, 5p12‐q11, 13q21.1, 18q24, 1q21‐q27, respectively. Moreover, real‐time quantitative RT‐PCR assays show that porcine Arf1‐6 are ubiquitous in all tissues examined, with the highest levels in the kidney and stomach and the lowest in muscle and the heart. This is the first report of molecular characterization of the Arf gene family in pigs. 相似文献
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J.‐J. Luo H.‐W. Song B. Zhang L.‐L. Li Y.‐G. Chen Y. Peng L.‐Z. Wu J.‐X. Fan J.‐S. Zhan 《Journal of animal physiology and animal nutrition》2014,98(3):517-521
To clone adiponectin (ADPN) gene from Shaziling porcine adipocyte and construct its eukaryotic expression vector, total RNA was extracted from subcutaneous fatty tissue. One pair of specific primers was designed by Primer 5.0 software according to the sequence of ADPN gene of porcine available in GenBank. The ADPN gene was amplified by PCR from cDNA and cloned into pMD18‐T vector to construct recombinant clonal vector pMD‐ADPN, sequenced and analysed. A recombinant expression plasmid pPICZaA‐ADPN was constructed by subcloning the cloned ADPN gene into the linearized pPICZaA vector. Then, the plasmid pPICZaA‐ADPN was expressed in Pichia pastoris (GS115) by electrotransformation. Western blot and Bradford analysis were used to determine the target protein induced by methanol. Results showed that the genome size of ADPN was 732 bp and encoded 244 amino acid, the nucleotide sequence of ADPN shared 100% identity with that of porcine available in GenBank. Western blot and Bradford analysis showed that the recombinant ADPN was expressed in GS115 correctly and has certain immune activity. The expression level of ADPN was 28.5 μg/ml. In conclusion, the recombinant ADPN could express in eukaryotic expression vector pPICZaA‐ADPN constructed in this study effectively. 相似文献
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XU Li-pu WANG Xiao-liang ZHANG Wen PAN Yong JING Hong-li WANG Jing-bo WANG Shu CAO Huan WANG Peng NA Li-hai JIANG Yu-lin 《中国畜牧兽医》2017,44(2):613-618
An acute infectious diseases occurred in a koi farm in Fangshan district, Beijing, and it resulted in mortalities of more than 50%.The main symptoms of sick koi were gills necrosis, kidney erosion and edema,which were similar to the clinical signs of koi herpes virus disease (KHVD).But PCR tests showed negative results for KHV. For further diagnosis, bacterial cultures, transmission electron microscopy studies, virus isolation and PCR tests were used. Electron microscopic observation revealed pox virus particles having a size of about 200 nm×400 nm in the kidney. 548 and 180 bp fragments were amplified from organs of sick koi by PCR method using specific primers of carp edema virus (CEV). The fragments were sequenced and analysed. The results showed that they were shared 100% nucleotide identity with CEV-H504. All the results indicated that this disease was carp edema virus disease, caused by a kind of pox virus, CEV. This was the first report on the CEV of cultured koi in China. 相似文献
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Jianpeng Gao Mingxing Tian Yanqing Bao Peng Li Jiameng Liu Chan Ding Shaohui Wang Tao Li Shengqing Yu 《Veterinary research》2016,47(1)
Brucellosis, caused by a facultative intracellular pathogen Brucella, is one of the most prevalent zoonosis worldwide. Host infection relies on several uncanonical virulence factors. A recent research hotpot is the links between carbon metabolism and bacterial virulence. In this study, we found that a carbon metabolism-related pyruvate kinase (Pyk) encoded by pyk gene (locus tag BAB_RS24320) was associated with Brucella virulence. Determination of bacterial growth curves and resistance to environmental stress factors showed that Pyk plays an important role in B. abortus growth, especially under the conditions of nutrition deprivation, and resistance to oxidative stress. Additionally, cell infection assay showed that Pyk is necessary for B. abortus survival and evading fusion with lysosomes within RAW264.7 cells. Moreover, animal experiments exhibited that the Pyk deletion significantly reduced B. abortus virulence in a mouse infection model. Our results elucidated the role of the Pyk in B. abortus virulence and provided information for further investigation of Brucella virulence associated carbon metabolism. 相似文献
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