Cloning and expression of canine interferon-alpha genes in Escherichia coli   总被引：1，自引：0，他引：1  

Taira O  Watanugi I  Hagiwara Y  Takahashi M  Arai S  Sato H  Maehara N 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(10):1059-1062

We cloned five new subtypes of cDNA encoding canine interferon-alpha (CaIFN-alpha) from a canine epithelial cell line. CaIFN-alphas were divided into two groups by amino acid sequences and a molecular phylogenic tree. Two subtypes of them were expressed in Escherichia coli, and IFN proteins were purified. Recombinant CaIFN-alphas were highly species-specific and showed antiviral activity against Vesicular stomatitis New Jersey virus and canine adenovirus-1 , but not against canine herpesvirus-1.  相似文献   

Vaccine efficacy of a cell lysate with recombinant baculovirus-expressed feline infectious peritonitis (FIP) virus nucleocapsid protein against progression of FIP     

Hohdatsu T  Yamato H  Ohkawa T  Kaneko M  Motokawa K  Kusuhara H  Kaneshima T  Arai S  Koyama H 《Veterinary microbiology》2003,97(1-2):31-44

The Type II feline infectious peritonitis virus (FIPV) infection of feline macrophages is enhanced by a monoclonal antibody (MAb) to the S protein of FIPV. This antibody-dependent enhancement (ADE) activity increased with the MAb that showed a neutralizing activity with feline kidney cells, suggesting that there was a distinct correlation between ADE activity and the neutralizing activity. The close association between enhancing and neutralizing epitopes is an obstacle to developing a vaccine containing only neutralizing epitopes without enhancing epitopes. In this study, we immunized cats with cell lysate with recombinant baculovirus-expressed N protein of the Type I FIPV strain KU-2 with an adjuvant and investigated its preventive effect on the progression of FIP. Cats immunized with this vaccine produced antibodies against FIPV virion-derived N protein but did not produce virus-neutralizing antibodies. A delayed type hypersensitivity skin response to N protein was observed in these vaccinated cats, showing that cell mediated immunity against the FIPV antigen was induced. When these vaccinated cats were challenged with a high dose of heterologous FIPV, the survival rate was 75% (6/8), while the survival rate in the control group immunized with SF-9 cell-derived antigen was 12.5% (1/8). This study showed that immunization with the cell lysate with baculovirus-expressed N protein was effective in preventing the progression of FIP without inducing ADE of FIPV infection in cats.  相似文献   

Polygalacturonase S31PG1 from <Emphasis Type="Italic">Geotrichum candidum</Emphasis> citrus race S31 expressed in <Emphasis Type="Italic">Schizosaccharomyces pombe</Emphasis> versus S31PG2 regarding soft rot on lemon fruit     

Masayuki?Nakamura  Hisashi?IwaiEmail author  Kei?Arai 《Journal of General Plant Pathology》2003,69(5):283-291

Gene S31pg1, which encodes a polygalacturonase (PG), was previously isolated from citrus race S31 of Geotrichum candidum, the causal agent of citrus sour rot. We have now isolated and sequenced an additional PG gene, S31pg2, with 95% identity to S31pg1 in the mature proteins. To evaluate the contribution of the two PG genes in the development of citrus sour rot, each gene was expressed in the fission yeast Schizosaccharomyces pombe. Both genes conferred PG activity to the yeast. Crude enzyme solutions containing S31PG1 severely degraded the albedo tissue of lemon peel, but those containing S31PG2 did not. Concentrated crude S31PG1 solutions also caused soft rot on lemon fruit, indicating that not S31PG2 but S31PG1 is an important pathogenicity factor in citrus sour rot. Next, the protopectinase (PP) activity of each PG was measured. Although S31PG1 and S31PG2 are highly homologous, S31PG1 had high PP activity, whereas S31PG2 had much lower activity. PG from G. candidum noncitrus race S63 (nonpathogenic to citrus fruits) was also assayed but did not have any PP activity at all. These results suggest that the different PP activities of the PGs are a key to the pathogenicity of G. candidum to lemon fruit.  相似文献   

First Report of <Emphasis Type="Italic">Pepper mottle virus</Emphasis> on <Emphasis Type="Italic">Capsicum annuum</Emphasis> in Japan     

Yoshihiro?OgawaEmail author  Kyoji?Hagiwara  Hisashi?Iwai  Shoichi?Izumi  Kei?Arai 《Journal of General Plant Pathology》2003,69(5):348-350

Pepper mottle virus, genus Potyvirus, was first identified in Japan based on particle morphology, host range, aphid transmission, and molecular classification using the nucleotide sequence of the coat protein gene and 3-untranslated region.  相似文献   

Nitrate reductase,a nitric oxide-producing enzyme: induction by pathogen signals   总被引：8，自引：1，他引：7  

Ayako?Yamamoto  Shinpei?Katou  Hirofumi?Yoshioka  Noriyuki?Doke  Kazuhito?KawakitaEmail author 《Journal of General Plant Pathology》2003,69(4):218-229

  相似文献   

Anthracnose of Sansevieria trifasciata caused by Colletotrichum sansevieriae sp. nov.     

Masayuki Nakamura  Masahiro Ohzono  Hisashi Iwai  Kei Arai 《Journal of General Plant Pathology》2006,72(4):253-256

A Colletotrichum sp. was isolated from water-soaked lesions on sansevieria (Sansevieria trifasciata Prain cv. Laurentii) in Japan. Classifying the species only from the morphology of the fungus was difficult; therefore, host range was tested and the ribosomal DNA ITS2 region was phylogenetically analyzed. The fungus was pathogenic only on sansevieria among 20 test plants belonging to 11 families. In a phylogenetic analysis with the neighbor-joining method, the two isolates used formed a single-isolate clade. The fungus is thus proposed to be a new species, Colletotrichum sansevieriae. This report is the first of anthracnose on sansevieria.  相似文献   

The Activity Ratio of the Cytosolic MDH/LDH and the Isoenzyme Pattern of LDH in the Peripheral Leukocytes of Dogs,Cats and Rabbits     

Washizu T  Nakamura M  Izawa N  Suzuki E  Tsuruno S  Washizu M  Nakajo S  Arai T 《Veterinary research communications》2002,26(5):341-346

The activities of malate dehydrogenase (MDH) and lactate dehydrogenase (LDH) and the pattern of the isoenzymes of LDH were determined in the peripheral blood leukocytes of dogs, rabbits and cats. Rabbits had significantly higher plasma glucose concentrations than dogs or cats. Feline leukocytes showed higher LDH and lower MDH activities than canine or rabbit leukocytes. The M/L ratio, defined as the MDH activity divided by the LDH activity in cytosolic fractions, was considered to be a good indicator with which to evaluate the metabolic state in animal tissues. The M/L ratio was highest in canine and lowest in feline leukocytes. LDH-2 and LDH-3 isoenzymes were dominant in canine leukocytes. LDH-1 and LDH-2 were dominant in rabbit leukocytes, whereas LDH-5 was dominant in feline leukocytes. It was evident that there were significant differences in energy metabolism between the leukocytes of dogs, rabbits and cats.  相似文献   

Expression of desmosomal proteins in rat keratinocytes during in vitro differentiation     

Mochizuki R  Kamiyama M  Arai KY  Arai K  Uehara K 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(2):123-127

The keratinocyte, the major component of the epidermis, expresses several proteins that characterize the keratinization during the differentiation. Proliferation and differentiation of cultured human keratinocytes are known to be regulated by the Ca2+ concentration in the culture medium. However, informations about the rat keratinocyte are relatively limited and their physiology is still an open question. To elucidate the characteristics of the rat keratinocyte, we established rat keratinocyte culture system and examined effects of extracellular calcium concentration on the expression of differentiation-related proteins. Keratinocytes were isolated from the newborn rat skin with 0.25% trypsin, followed by separation with a Percoll density gradient. The separated cells were grown in MCDB 153 medium containing several growth factors and Ca(2+)-free fetal bovine serum, then stimulated with Ca2+. Immunoblotting demonstrated strong expression of beta1 integrin in unstimulated cells, suggesting that the primary culture of rat keratinocytes was successfully established. Expression of desmoglein and transglutaminase was increased by Ca2+ stimulation, whereas beta1 integrin expression was decreased in response to increasing concentrations of Ca2+. These observations indicate that cultured rat keratinocytes maintain the ability to differentiate in vitro, which is similar to that of the basal keratinocytes in the epidermis.  相似文献   

Changes in hepatic enzyme activities in transgenic mice carrying human prototype c-Ha-ras gene treated with diethylnitrosamine     

Arai T  Ogawa T  Nakamura M  Hosoya M  Ohnishi Y 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(11):1065-1067

Activities of malate dehydrogenase (MDH) and aspartate aminotransferase in the malate-aspartate shuttle were significantly increased in the cytosolic fractions of livers with early neoplastic symptoms such as swelling and discoloration in transgenic mice carrying the prototype human c-Ha-ras gene, Tg-rasH2 mice, which were administered with diethylnitrosamine (DEN) as a carcinogenic chemical at a dose of 200 mg/kg body weight. Cytosolic MDH/LDH (lactate dehydrogenase) (ML) ratio increased significantly and was considered to be a useful marker to characterize the energy metabolism at early neoplastic stage in livers of the Tg-rasH2 mice.  相似文献   

Plasma glucose,mannose, and non-esterified fatty acid concentrations in layer-type chickens     

T. Sato  S. Soeda  Y. Ohta  K. Katayama  T. Arai  H. Tazaki 《British poultry science》2008,49(6):731-735

1. The oral administration of glucose or dietary glucose reduces fasting plasma mannose concentrations in mammals. On the other hand, there have been no reports on plasma mannose levels in birds. We have analysed chicken plasma mannose and glucose by an original high-performance liquid chromatography (HPLC) method, together with plasma non-esterified fatty acid (NEFA) concentrations in chickens.

2. Plasma glucose concentrations of chickens did not differ among three different age groups (0, 18 and 150 d). However, the plasma mannose concentrations of chicks at the age of 0 d were higher than those of chickens at the ages of 18 and 150 d.

3. At the age of 18 and 150 d, plasma glucose concentrations were elevated and plasma mannose and NEFA concentrations were decreased after regular feeding, compared to fasting levels.  相似文献