犏牛和牦牛ESCO2基因的序列特征及其在睾丸中的表达对比分析     

闵星宇  杨丽雪  杨满珍  胡宇磊  于海玲  杨璐瑜  李键  熊显荣 《畜牧兽医学报》2021,52(11):3126-3136

旨在克隆犏牛和牦牛姐妹染色单体内聚建立蛋白2（establishment of sister chromatid cohesion N-acetyltransferase 2,ESCO2）基因,并分析其在不同发育阶段睾丸中的表达与定位,为进一步解析ESCO2在减数分裂过程中的作用机制提供理论依据。本研究以健康雄性犏牛及牦牛为试验动物,根据年龄分为胎牛组（5~6月龄）、幼年组（1~2岁）和成年组（3~4岁）,每组各3头。通过RT-PCR技术克隆犏牛和牦牛ESCO2基因并进行生物信息学分析,采用实时荧光定量PCR （quantitative real-time PCR,qRT-PCR）检测ESCO2基因在犏牛不同组织中的表达谱,比较分析ESCO2在犏牛和牦牛不同时期睾丸中的表达规律,利用免疫组织化学（immunohistochemistry,IHC）技术检测ESCO2蛋白的细胞定位和表达差异。结果显示,犏牛ESCO2基因（GenBank登录号：MW198470） CDS区为1 833 bp,编码610个氨基酸,与牦牛相比,犏牛ESCO2序列第301~319位多19个氨基酸,另有3个氨基酸突变;犏牛ESCO2蛋白序列与黄牛的同源性高于其他哺乳动物;ESCO2可能与SMC3、SMC1A、PDS5A、PDS5B、STAG2等蛋白相互作用,互作蛋白功能与姐妹染色单体凝聚、减数分裂细胞周期、DNA修复、细胞分裂和染色体重构等生物学过程相关。ESCO2在犏牛各组织中均有表达,但在睾丸中的相对表达水平显著高于其它组织（P<0.05）;在犏牛睾丸中的表达随年龄增长呈上升趋势,幼年和成年时期犏牛睾丸中ESCO2的表达显著低于同时期牦牛（P<0.05）;IHC染色结果发现,雄性犏牛减数分裂阻滞于初级精母细胞,ESCO2蛋白在犏牛初级精母细胞中无表达并与牦牛存在差异。本研究结果表明,犏牛与牦牛的ESCO2基因、蛋白序列差异较大,且在睾丸发育过程中的表达模式差异显著,这可能是引起雄性犏牛减数分裂阻滞及不育的原因之一,其具体作用机制有待进一步研究。  相似文献   

猪ADAR2基因全长cDNA克隆、序列特征及表达模式分析     

张跃博  王立刚  侯欣华  刘欣  颜华  张龙超  王立贤 《畜牧兽医学报》2021,52(3):620-629

旨在克隆猪作用于RNA的腺苷脱氨酶2基因（ADAR2）全长cDNA序列,同时对该基因在猪不同组织中的表达规律进行探索。利用RACE （rapid-amplification of cDNA ends）对大白猪ADAR2基因mRNA全长序列进行克隆,并进行生物信息学分析;用荧光定量PCR方法检测35日龄大白猪心、肝、肺、肾、脾、脑、小肠、背最长肌和背部脂肪9种组织中ADAR2的表达水平。结果表明,猪ADAR2基因cDNA全长6 305 bp,共包含12个外显子,编码704个氨基酸,与人、黑猩猩、猕猴、长臂猿、黄牛、山羊和绵羊的CDS区核酸序列和氨基酸序列的一致性均在84%以上。该基因编码的蛋白含有2个双链RNA结合基序和一个脱氨酶结构域。猪ADAR2在检测的各组织中均表达,其中在肺中的表达量最高。综上所述,本研究成功克隆了猪ADAR2基因全长cDNA序列,并且发现其在猪体内广泛表达,为深入研究ADAR2的功能奠定了良好的基础。  相似文献   

Genomic characterisation of bovine papillomavirus types 1 and 2 identified in equine sarcoids in Japan     

Nanako Yamashita-Kawanishi  James K. Chambers  Kazuyuki Uchida  Yumiko Tobari  Hisashi Yoshimura  Masami Yamamoto  Norio Yumoto  Hiroshi Aoki  Katsuaki Sugiura  Tohru Higuchi  Shigeaki Saito  Takeshi Haga 《Equine veterinary journal》2021,53(6):1199-1209

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Identification of the core motif of the BRCA2 C-terminal RAD51-binding domain by comparing canine and human BRCA2     

Yasunaga YOSHIKAWA  Masami MORIMATSU  Kazuhiko OCHIAI  Toshina ISHIGURO-OONUMA  Ryo MORIOKA  Kento OKUDA  Koichi ORINO 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(5):759

Mammary tumors are the most common tumors in women and non-spayed female dogs. One of the reasons for mammary tumors is mutations of the tumor suppressor gene, BRCA2. BRCA2 participates in homologous recombination repair by interacting with the RAD51 recombinase. BRCA2 has two RAD51-binding domains, consisting of BRC repeats and the C-terminal RAD51-binding domain, respectively. Although several studies have addressed the function of the C-terminal RAD51-binding domain of human BRCA2, the amino acid sequences required for the RAD51-interaction activity remain unclear. In this study, the C-terminal RAD51-binding domains of canine and human BRCA2 were compared; the canine domain displayed a weaker interaction with RAD51. This difference was attributed to the C-terminal portion of the domain via a comparison between canine and human domains. Furthermore, peptides shorter than those previously reported displayed RAD51-interacting activity, and a core motif of this domain consisting of 25 amino acids was identified. Since a mutation (S3323N) was reported in the core motif of this domain, the effect of this mutation was evaluated. The mutant exhibited similar RAD51-binding activity as that of the wild-type protein, suggesting that the mutation was functionally neutral. These data suggested that the C-terminal portion of the BRCA2 C-terminal RAD51-binding domain influenced its RAD51-interaction activity, and a minimum core motif of 25 amino acids was identified in this domain. These data may help clarify BRCA2 function, as well as the tumorigenic effects of BRCA2 mutation.  相似文献   

1株猫杯状病毒的分离鉴定及其ORF2序列的遗传演化分析     

李鑫  杨德全  葛菲菲  沈海潇  刘健  鞠厚斌  王建  赵洪进 《畜牧与兽医》2021,(2):92-96

为了解猫杯状病毒形态特征及遗传演化情况,采用F81细胞从患病宠物猫的鼻拭子样品中分离获得1株猫杯状病毒(feline calicivirus,FCV),命名为SH1。经电镜观察,病毒粒子呈球形,无囊膜,符合FCV的形态特征。采用RT-PCR方法扩增了该毒株的全基因组,并进行了序列测定和衣壳蛋白基因(ORF2)序列的分析。结果显示:分离株与国内外参考株的ORF2序列的核苷酸和氨基酸同源性分别为74.1%~79.7%和84.5%~90.9%;ORF2基因的遗传演化分析显示,30株FCV毒株形成两大分支,即基因群Ⅰ和Ⅱ,分离株属于基因群Ⅰ;进一步分析发现,基因群Ⅰ和Ⅱ主要在377、539和557氨基酸位点存在差异,基因群Ⅰ和Ⅱ分别为N、A、G和K、V、S。研究结果为FCV感染的防控提供科学依据。  相似文献   

Single oral β-cryptoxanthin administration increases its serum concentration and enhances peripheral blood neutrophil function in Holstein cattle     

Atsushi KIMURA  Yo-Han KIM  Kazuyoshi HASHIZUME  Akira ITO  Katsuyuki MUKAI  Keiichiro KIZAKI  Shigeru SATO 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(5):829

We investigated the effect of oral administration of β-cryptoxanthin (β-CRX) on its serum concentration and peripheral neutrophil functions by the chemiluminescence (CL) response in Holstein cattle. A single oral administration of β-CRX was performed for serum β-CRX concentration (0, 0.05, 0.1, or 0.2 mg/kg body weight [BW]) and for peak CL response of peripheral neutrophils (0.2 mg/kg BW). The serum β-CRX concentration was peaked on 2 days after, similar to peak CL response on 3 days after β-CRX administration. Therefore, a single oral administration of β-CRX (0.2 mg/kg BW) induces higher serum concentration and concurrently enhances bactericidal ability of peripheral neutrophils in Holstein cattle.  相似文献   

Molecular characterization of Ascaridia galli from Bangladesh and development of a PCR method for distinguishing A. galli from Heterakis spp.     

Peru Gopal BISWAS  Yuma OHARI  Uday Kumar MOHANTA  Tadashi ITAGAKI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(4):666

We analyzed the nuclear ribosomal internal transcribed spacer (ITS) 1 and ITS2 sequences for Bangladesh isolates of Ascaridia galli, and we determined that the sequences were unreliable as molecular markers for distinguishing A. galli from other Ascaridia species, because the sequences showed high identity with that of A. columbae. However, the ITS1 sequences were available for designing PCR primers distinguishable between Ascaridia galli and Heterakis spp. Bangladesh isolates of A. galli constituted a monophyletic clade along with other geographical isolates in the cytochrome c oxidase subunit I (COI) phylogenetic tree, however, we could not clarify the phylogenetic relationships between A. galli and other Ascaridia spp., because their available sequences in GenBank were very few. The developed PCR method using DNA from A. galli and Heterakis spp. eggs would enable differential diagnosis of the individual infections in the future.  相似文献   

Immunohistochemical expression of HER - 2 and Ki - 67 in granulosa cell tumor in bitches     

Ana Carina Heil da Silva Matos  Angélica Consalter  Bárbara Paula dos Santos Batista  Ana Beatriz Monteiro Fonseca  Ana Maria Reis Ferreira  Juliana da Silva Leite 《Reproduction in domestic animals》2021,56(4):667-672

Granulosa cell tumour, an ovarian neoplasm of stromal origin, is an important tumour related to oestrogenic dominance syndrome and cystic endometrial hyperplasia–pyometra complex. In order to analyse ovarian tumour´s malignant potential, immunohistochemical markers can be used, such as anti-HER2 and anti-Ki-67. The aim of this study was to evaluate the expression of immunohistochemical markers HER-2 and Ki-67 in granulosa cell tumour from bitches´ ovaries. In HER-2 immunomarker analysis using the HercepTest^® method, most tumours were classified as 2+ (moderate labelling). Concerning Ki-67 immunomarker, only one case was described as having a high proliferative index. An association was found between immunostained cell percentage by anti-HER-2 antibodies and high pleomorphism, represented by the pattern of follicular/trabecular tumour arrangement. There was no correlation between anti-Ki-67 and anti-HER-2 antibody immunostaining intensities, probably due to only one case with a high Ki-67 index. With an effective protocol for HER-2 and Ki-67 immunohistochemical identification in granulosa cell tumours in bitches, it was possible to characterize this neoplasm proliferation profile.  相似文献   

The influence of progesterone and prostaglandin F2α on decorin and the adhesion of caruncular epithelial cells of bovine placenta at early-mid pregnancy—Part II     

Monika Jamioł  Jacek Wawrzykowski  Marta Kankofer 《Reproduction in domestic animals》2021,56(7):1040-1049

One of the most important processes determining the proper course of gestation and its physiological termination in cows is the adhesion of epithelial cells allowing for direct contact of maternal and foetal parts of the placenta. Throughout pregnancy, placental cells are under strict hormonal control, which among others regulates the concentration and activity of specific proteins participating in the extracellular matrix remodelling of foetal membranes. The aim of the study was to evaluate the influence of progesterone and prostaglandin F_2α on the adhesion of epithelial cells at early-mid pregnancy in cows. Additionally, the impact of selected hormones on anti-adhesive properties of decorin was evaluated. Caruncular epithelial cells were isolated from healthy cows during pregnancy, immediately after slaughter. Primary cell cultures derived from the 2nd and 4th month of gestation were used in the experiments. The viability of cells was assessed by MTT assay. The adhesion of cells to fibronectin was measured spectrophotometrically. The activity of metalloproteinases was confirmed by the metalloproteinase assay. Progesterone (10^–5 and 10^–7 mol/L) and prostaglandin F_2α (10^–4_, 10^–5 and 10^–7 mol/L) increased the viability of bovine caruncular epithelial cells in the 2nd month of pregnancy. The treatment with prostaglandin F_2α significantly reduced the number of adherent cells from the 2nd month of gestation at the doses of 10^–4 and 10^–5 mol/L. Both progesterone and prostaglandin F_2α were shown to have an effect of decorin resulting in both a decrease in metalloproteinase activity and an increase in adhesion of cells to fibronectin.  相似文献   

培肥措施对旱地农田土壤CO2排放和碳库管理指数的影响     

王晓娇  蔡立群  齐鹏  王雅芝  陈晓龙  武均  张仁陟 《草业学报》2021,30(2):32-45

阐明长期有机物料施肥下土壤CO₂排放特征及其影响机制以及碳库管理指数对黄土高原旱作农业区固碳减排及施肥模式选择的影响尤为重要。基于2012年设置在陇中黄土高原旱作区的长期定位试验,通过不施肥（CK）、氮肥（NF）、有机肥（OM）、秸秆（ST）、有机肥结合无机肥（OMNF）5个处理,测定并计算了2018年不同施肥措施下全年土壤CO₂排放、作物碳排放效率和碳库管理指数的变化,并运用结构方程模型分析了0~30 cm土壤温度、水分、微生物量碳氮、易氧化有机碳、蔗糖酶、脲酶与土壤CO₂排放速率的关系。结果表明：1）与不施肥相比,秸秆、有机结合无机肥和有机肥处理使生育期土壤CO₂排放平均速率提高了42.72%、30.82%和29.79%,秸秆、有机肥处理分别使生育期土壤CO₂排放量显著提高36.35%、32.45%（P<0.05）,有机结合无机肥处理使碳排放效率显著降低41.10%（P<0.05）;2）有机物料处理均能显著提高0~5 cm土层易氧化有机碳、微生物量碳氮、蔗糖酶活性和碳库管理指数,相比不施肥和氮肥处理,有机结合无机肥处理分别使0~30 cm土壤碳库管理指数提高127.41%,99.33%（P<0.05）;3）结构方程模型表明,环境因子对土壤CO₂排放速率的总解释度为53%,对土壤CO₂排放速率总效应较大的影响因素包括土壤温度（2.36）、微生物量碳（1.59）和土壤水分（1.18）,且均间接地影响着土壤CO₂排放速率,土壤温度促进了微生物量碳和蔗糖酶活性的提高,微生物量碳促进了微生物量氮和易氧化有机碳的增加。综合来看,有机结合无机肥处理可以提升土壤碳库管理指数,保持微生物活性,增加作物产量,降低土壤碳排放效率,是陇中黄土高原旱作农业区比较适宜的农田培肥措施。  相似文献