传染性支气管炎病毒NP基因在大肠杆菌中的可溶性表达     

郝春丽  王泽霖  王新卫  姚慧霞  周欣 《安徽农业科学》2007,35(34):11022-11024

[目的]生产具有生物活性的、可用于临床检测的NP融合蛋白抗原,建立可用于临床检测IB抗体诊断的试剂盒。[方法]利用RT-PCR技术扩增传染性支气管炎病毒M41株的NP基因,将之克隆并重组于原核表达载体pET-28a,经PCR、酶切鉴定及序列分析构建重组表达载体pET-NP,研究NP基因在大肠杆菌中的可溶性表达。[结果]将重组载体pET-NP转化表达菌BL21,获得阳性重组菌pET-NP-BL。阳性重组子经IPTG诱导后,目的基因NP获得了高效表达。表达产物经过SDS-PAGE与Western blot检测表明,NP分子量约为49 kD,且以可溶性蛋白形式存在;表达产物可与特异性传染性支气管炎病毒抗血清发生免疫反应。表达产物可以用作检测IB抗体的抗原。[结论]该融合蛋白可用于生产检测IB的诊断试剂,生物安全性高,生产方便。但尚需建立适当的检测方法,进一步研究其实际诊断价值。同时也可以研制新型IB重组亚单位疫苗,进一步研究其刺激动物机体产生的细胞与体液免疫,深入研究新功能,筛选新的抗原表位,为生产新型IB基因工程疫苗奠定基础。  相似文献   

Role of miR-219 and TGFBR2 in pathogenesis of renal fibrosis     

LI Shuang-qing  YING Jun  XU Xu-chun 《园艺学报》2019,35(4):703-709

AIM:To study the role of microRNA-219 (miR-219) in regulation of transforming growth factor-β receptor type 2 (TGFBR2) in renal fibrosis. METHODS:The renal fibrosis patients (n=70) were selected in this stu-dy, and 20 cases of healthy people were selected as control group. RT-qPCR was used to detect the expression of miR-219 in the serum of the patients with renal fibrosis and control group, and the expression of miR-219 in NRK49F cells after stimulation with angiotensin Ⅱ(AngⅡ) was detected. The protein expression of α-smooth muscle actin (α-SMA) in the NRK49F cells transfected with miR-219 mimics after stimulation with AngⅡ was determined by Western blot. The potential target gene TGFBR2 of miR-219 was screened and verified by the method of luciferase reporter gene. RT-qPCR and Western blot were used to detected the effect of miR-219 mimics on the expression of TGFBR2 at mRNA and protein levels, and the mRNA expression of α-SMA, connective tissue growth factor (CTGF), type I collagen α1 (COL1A1) and COL3A1 in the NRK49F cells was also detected, respectively. The unilateral ureteral occlusion (UUO) mouse model was established and the expression of miR-219 in the renal tissue was monitored. The morphological change of renal fibrosis was observed in the UUO mice after injection of miR-219, and the mRNA expression levels of COL1A1 and COL3A1 were detected. RESULTS:The expression level of miR-219 in the patients with renal fibrosis was significantly lower than that in control group, and the expression of miR-219 in the UUO mice was decreased significantly (P<0.01). The expression level of miR-219 was significantly decreased in the NRK49F cells after AngⅡ stimulation, and miR-219 mimics inhibited the protein expression of α-SMA(P<0.01). miR-219 mimics had a targeted regulatory effect on TGFBR2 gene, which inhibited the mRNA and protein expression of TGFBR2. miR-219 mimics inhibited the mRNA expression of α-SMA, CTGF, COL1A1 and COL3A1. miR-219 also down-regulated the mRNA expression of COL1A1 and COL3A1 in the UUO mice and inhibited the process of renal fibrosis. CONCLUSION:miR-219 inhibits the development of renal fibrosis by inhibiting the expression of TGFBR2, which may become a new target for the diagnosis and treatment of renal fibrosis.  相似文献   

中国假霜霉属分类的研究     

陶家凤  秦芸 《四川农业大学学报》1986,(1)

本文报告对我国5种假霜霉的研究结果,其中荨麻假霜霉(Pseudoperonosporaurticae)是在中国的首次记录。标本保存于四川雅安四川农业大学。  相似文献   

小麦品种对土传小麦黄色花叶病毒病抗性遗传的初步研究   总被引：1，自引：0，他引：1  

秦家忠  黎中明  陶家凤  秦芸 《四川农业大学学报》1986,(1)

本试验通过杂交、回交和抗病性鉴定等方法,测定了14个组合的F_1代对土传小麦黄色花叶病毒病的抗性遗传。根据双亲正反杂交表现型一致的结果,可以确定控制小麦品种对该病毒病的抗性为细胞核遗传。通过对F_1、F_2、BC_1及BC_2各代群体抗病株和感病株分离比值的分析,初步确定有关土传小麦黄色花叶病毒病的抗性遗传可能受两对致病显性基因(S_1、S_2),和一对抑制基因(I)所控制,作者并对这一问题与前人工作作了比较和讨论。  相似文献   

四川冕宁县缺硒饲粮对雏鸡的影响及不同品种雏鸡适宜补硒量的研究     

刘永钢  张光柱  王康宁  端木道  杨凤 《四川农业大学学报》1986,(1)

采用四川严重缺只硒地区冕宁县实用雏鸡饲粮饲喂泸宁鸡(当地地方鸡)、白洛克和来杭三个品种共682只雏鸡,作了两个试验,结果表明:谷实类缺硒饲粮(含硒0.0051ppm)引起雏鸡严重营养性胰腺萎缩和纤维化;泸宁鸡和白洛克缺硒死亡率分别为83％和63％,添加维生素E可起到轻微缓解作用(缺硒死亡率52％);补硒显著促进生长和改善饲料利用率。饲粮中添加动物性饲料(使基础日粮含硒0.1018ppm)后雏鸡未出现明显缺硒病变,但补硒显著提高谷胱甘肽过氧化物酶(GSH-px活力、组织硒含量和生长速度。雏鸡对缺硒的敏感程度、组织硒含量和GSH-px活力存在明显品种差异。以GSH-px活力和组织硒含量作为评定标准结合对生长速度的考察表明:对于生长较快的泸宁鸡和白洛克,基础饲粮补加0.20ppm硒是适宜的,而对于蛋用型来杭鸡则以添加0.10ppm效果较好。  相似文献   

抗A型禽流感病毒核蛋白特异性单克隆抗体研究     

李娜  秦爱建  邵红霞  金文杰  刘岳龙 《农业科学与技术》2008,9(1):60-63

禽流感（AI）是由A型禽流感病毒（AIV）引起的一种发生于禽类的病毒性传染病。笔者以杂交瘤技术研制抗AIV共同抗原的特异单克隆抗体,旨在建立一种双抗体夹心ELISA方法快速检测AIV,以便为A型AIV的快速诊断技术的研究提供理论依据。  相似文献   

猪流感病毒NP基因的克隆及杆状病毒转移载体的构建     

赵朴  郑玉姝  贾贝贝  乔传玲  陈化兰  刘兴友  李海燕 《西南农业学报》2009,22(5)

本研究通过RT-PCR扩增了猪流感病毒A/ Swine/ Inner Mongolia/ 547/ 01 (H3N2)的NP基因,并将其克隆入pMD 18-T载体.重组pMD 18-T经Sal I 和 EcoR I酶切后得到的NP基因插入pMelBacB载体的Sal I/EcoRI位点.PCR和限制性内切酶分析鉴定表明成功构建了pMelBacB-NP转移载体.这为开发NP诊断抗原和亚单位疫苗奠定了基础.  相似文献   

Enhanced kernel set promoted by synchronous pollination determines a tradeoff between kernel number and kernel weight in temperate maize hybrids     

Martín Uribelarrea  Jorgelina Cárcova  Lucas Borrás  María E. Otegui 《Field Crops Research》2008

Maize (Zea mays L.) grain yield is strongly related to the number of harvested kernels, where kernel number can be increased by synchronously pollinating silks rather than allowing them to be progressively pollinated as they naturally appear from the husks. However, there is scarce evidence on how this practice affects kernel weight (KW) and plant grain yield (PGY), and no report exists on its effects when combined with treatments aimed to reduce apical dominance, like male sterility and detasseling. Field experiments were conducted in two growing seasons (Exp₁ and Exp₂) using two hybrids, cropped at contrasting stand densities (3 and 9 plants per m²) and including (i) male-fertile and male-sterile versions, (ii) tasseled and detasseled plants, and (iii) natural (NP) and synchronous pollination (SP; pollen added manually to ears bagged 5 days after initial silking) systems. Tassel growth of sterile and fertile versions was also evaluated in a separate experiment (Exp₃). Detasseling increased the number of ears per plant reaching silking (P < 0.001) of NP plants, but this beneficial effect of reduced apical dominance did not improve kernel number per plant (KNP) or PGY. Similarly, the early arrest of anther growth in male-sterile plants had no clear benefit on KNP. In contrast, KNP was enhanced by synchronous pollination (range between −13% and +71%; average of +15.4% in Exp₁ and +3.9% in Exp₂). However, this pollination system promoted a decreased in KW (range between −30% and +4%; average of −11.8% in Exp₁ and −7.8 in Exp₂) such that the treatment had no effect on PGY (range between −19% and +37%; average of +1% in Exp₁ and −4% in Exp₂). Because plant growth rate around flowering was not different between pollination treatments, assimilate availability per kernel was reduced from ovary fertilization onwards in synchronously pollinated plants when compared to open pollinated plants. This explains the reduced KW when increasing KNP by synchronous pollination. In summary, none of the imposed treatments allowed grain yield to be increased at the plant level.  相似文献   

抗A型禽流感病毒核蛋白特异性单克隆抗体研究   总被引：6，自引：4，他引：2  

李娜  秦爱建  邵红霞  金文杰  刘岳龙 《扬州大学学报(农业与生命科学版)》2007,28(1):9-12

利用禽流感H9亚型病毒(AIV-H9)免疫Balb/c小鼠,取其脾细胞与骨髓瘤细胞进行融合,经免疫荧光试验(IFA)检测,以研制抗禽流感病毒(AIV)单克隆抗体。结果获得了5株特异性抗AIV核蛋白(NP)的单克隆抗体细胞株,分别命名为AIV-NP-2C3、AIV-NP-6A5、AIV-NP-3H9、AIV-NP-7B4、AIV-NP-2H4。这5株单克隆抗体能与所有试验的AIV-H9病毒反应,Western blotting方法鉴定结果表明,单克隆抗体仅识别60 ku的蛋白抗原,而不与新城疫病毒、禽网状内皮组织增殖症病毒、传染性法氏囊病毒等反应。初步应用结果显示,以这些单克隆抗体建立的间接免疫荧光试验或ELISA方法可迅速检测出禽流感病毒,这些单克隆抗体在禽流感的预防监测中将发挥重要的作用。  相似文献   

玉米个体发育进程中控制叶片生长的基因效应差异性初探   总被引：1，自引：1，他引：0  

赖仲铭  杨克诚 《四川农业大学学报》1985,(2)

通过对玉米各主要生育时期叶片性状的基因效应及配合力方差分析结果表明;玉米个体发育不同时期的叶片性状(单株出叶数、绿叶面积,出叶速率、叶面积增长速率)所受的基因主导效应是有较大差异的。因此,如果我们沿用以往的研究方法,以终结产物来推源(估算、推论)供试性状的基因型或基因效应,显然只能表示该性状终结阶段的状况,在此前后作用于该性状的基因效应不可能尽相一致的。  相似文献