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41.
鸡Mx基因全长cDNA序列的克隆及分析 总被引:2,自引:0,他引:2
以Poly(I)-Poly(C)诱导鸡成纤维细胞Mx基因的表达,提取总RNA,RT-PCR扩增出全长Mx cDNA,扩增产物克隆入pMD19-T Simple载体中进行序列测定。结果表明:与GenBank公布的鸡Mx cDNA序列相比,其同源性达99.9%,为进一步研究鸡Mx基因的抗病毒活性和作用机理奠定了基础。 相似文献
42.
XIAO Gang ZHANG Zhen-qian LIU Rui-yang YIN Chang-fa WU Xian-meng TAN Tai-long GUAN Chun-yun 《中国农业科学(英文版)》2013,(6):962-970
Based on the sequence of a novel expressed sequence tag (EST), the full-length cDNA of 1 017 nucleotides was cloned from Brassica napus cv. Xiangyou 15 through rapid amplification of cDNA ends (RACE). The gene was designated as Bnhol34 (HQ585980), encoding a protein of 338 amino acids. BLAST analysis showed no high degree of sequence identity to any known gene. The calculated molecular weight of the Bnhol34 protein was 36.23 kDa, and the theoretical isoelectric point was 8.74. The Bnhol34 was also cloned from a high oleic acid mutant 854-1 through homologous cloning. There was no difference between the two Bnhol34 genes. Bnhol34 was localized in a tissue-specific manner in B. napus, and its expression level was about eight-fold greater in Xiangyou 15 seeds than in 854-1. The promoter region sequences of Bnhol34 were then isolated from Xiangyou 15 and 854-1, and a 93-bp deletion was found to occur in the Bnhol34 promoter region of 854-1. Three abscisic acid-responsive cis-elements (ABRE) were identified in the promoter region of Xiangyou 15. Real-time PCR analyses revealed that exogenous abscisic acid increased Bnhol34 expression by about four-fold in Xiangyou 15 seeds, yet did not change Bnhol34 expression in 854-1. It appeared that Bnhol34 might be abscisic acid insensitive in 854-1. 相似文献
43.
通过高校图书馆“一站式”导读系统的特点、设计思想及界面设计,阐述了本导读系统的功能以及实现该功能的关键技术。作为信息传播途径之一,导读系统的设计,提高了高校图书馆的服务质量和宣传方式。 相似文献
44.
We have established a simple diagnosis method for rice blast fungus resistant to MBI-D. This involves the preparation of PCR templates directly from the lesions in combination with primer-introduced restriction enzyme analysis PCR (PIRA-PCR). 相似文献
45.
The commercial LCx amplification assay, usually employed to detect the Myocobacterium tuberculosis complex in respiratory specimens, was evaluated by comparing the results it gave with those obtained using Löwenstein-Jensen solid medium and pathological findings on 55 lymph nodes from cattle with positive and 10 lymph nodes from cattle with negative skin tests for tuberculosis. Fifty-three cultures (51 and 2, respectively) were positive for M. bovis, while the results for the LCx assay and the histological method were positive in 48 (45, 3) and 24 (20, 4) samples, respectively. None of the samples from cattle from certified tuberculosis-free herds were positive by any of the procedures. The results obtained with the LCx assay, compared with the culture procedure, regarded as the gold standard among the diagnostic techniques, gave a specificity of 91.6% and sensitivity of 90.5%. Although the sensitivity of LCx was suboptimal, DNA of M. bovis was detected in 81.8% of the skin test-positive animals. Amplification techniques could provide a rapid and reasonably reliable tool for detecting bovine tuberculosis. 相似文献
46.
47.
旨在建立一种以重组酶聚合酶扩增技术(RPA)为基础的快速检测方法,用于小鹅瘟病毒的快速检测。小鹅瘟是一种常见的水禽传染病,严重危害我国养鹅业的健康发展。为了快速准确对小鹅瘟进行诊断,减少该病的危害,本研究以小鹅瘟病毒VP3基因保守片段为靶点,利用重组酶聚合酶扩增技术(RPA)建立了一种准确高效的小鹅瘟病毒RPA恒温快速检测方法,并对检测方法的灵敏度、特异性进行评价,并与传统PCR和传统RT-PCR方法进行比较。结果表明,该检测方法具有较高的灵敏度,可检测到10 copies/μL的病毒核酸;具有良好的特异性,只特异性地扩增鹅细小病毒,而与鹅副黏病毒、鹅源鸭瘟病毒、小鹅流行性感冒病毒、鹅副伤寒病毒、大肠杆菌和曲霉菌均未发生交叉反应;同时该方法重复性检测的变异系数低于6%,具有很好的重复性。阳性符合试验表明该检测方法与荧光定量PCR符合率为99%。该方法可以很好地应用于小鹅瘟的大规模临床样本检测,为小鹅瘟病毒的高通量检测和流行病学调查提供技术手段。 相似文献
48.
49.
4 香豆酸 :CoA连接酶 (4CL)是木质素生物合成过程中重要的酶 .该文利用简并寡核苷酸PCR法结合cDNA末端快速扩增PCR法直接获得了紫穗槐 4CLAcDNA全长序列 ,避免了复杂的构建、筛选cDNA文库的过程 .先用简并PCR得到了 4CLA1片段 ,又据此片段设计反向嵌套引物 ,用RACE方法获得未知的 5′和 3′端序列 .所获得的全长 4CLAcDNA ,编码 5 40个氨基酸 .氨基酸序列同源性分析表明4CLA是典型的 4CL蛋白 ,含有预计的AMP binding位点、催化反应区和保守的Cys . 相似文献
50.
Weidong SUN Xu YANG Hengbin QIU Di ZHANG Huanan WANG Jian HUANG Degui LIN 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(11):1541-1543
The BRCA1 gene plays an important role in the development of human breast
cancer, and recent research indicated that genetic variations of BRCA1 are also related to
canine mammary tumors (CMTs). Here, using rapid amplification of cDNA ends (RACE), we
cloned the 5′- and 3′-UTRs of BRCA1. By direct sequencing of the flanking sequences of the
5′- and 3′-UTRs of BRCA1, three previously unreported single-nucleotide polymorphisms
(SNPs) were identified, two (−1228T >C, −1173C >T) in the putative promoter regions
and one non-synonymous SNP (63449G >A) in exon 23. Compared with 16 normal samples, the
sequences from 34 CMTs suggested that SNP (−1173C >T) was associated with the
development of CMTs (odds ratio (OR)=2.57, 95% confidence interval (CI): 1.07–6.15). 相似文献