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三种PCR引物法制备生物素标记的HpLV衣壳蛋白基因cDNA探针检测效果的比较
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作者 韩盛 刘升学 +1 位作者 向本春 曹连莆 《石河子大学学报(自然科学版)》 CAS 2006年第4期415-418,共4页
采用正义、反义引物及正义和反义引物双链PCR法制备了生物素标记的HpLV衣壳蛋白基因的cDNA探针。检测结果表明,三种PCR引物法标记的cDNA探针显色灵敏度均可达到10pg/μL,对目标基因DNA的检测灵敏度可达90pg/μL,但反义引物PCR法标记探... 采用正义、反义引物及正义和反义引物双链PCR法制备了生物素标记的HpLV衣壳蛋白基因的cDNA探针。检测结果表明,三种PCR引物法标记的cDNA探针显色灵敏度均可达到10pg/μL,对目标基因DNA的检测灵敏度可达90pg/μL,但反义引物PCR法标记探针的检测效果要好于正义和反义引物双链PCR法制备的探针;在目标基因DNA浓度一定的情况下,探针的浓度对检测效果的影响并不大;正义、反义引物PCR法制备的探针可不经煮沸变性处理直接使用;用50ng/mL浓度的反义引物PCR法制备的生物素标记cDNA探针可检测到带毒啤酒花叶片和花瓣中的啤酒花潜隐病毒RNA。 展开更多
关键词 hplv 核酸杂交 生物素标记探针 cDNA检测效果
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N-羟基琥珀酰亚胺-α-萘乙酸酯的合成及其作为柱前衍生试剂在反相高效液相色谱中的应用
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作者 徐国良 王红 《化学试剂》 CAS CSCD 1999年第1期8-10,25,共4页
合成了N-羟基琥珀酰亚胺-α-萘乙酸酯(SINA)。研究了这一试剂在反相高效液相色谱中作为新的柱前衍生试剂,紫外可见检测,分离测定氨基酸的基本条件。衍生条件简单,方便。RP-HPLC分离了多种氨基酸,检测下限达pmo... 合成了N-羟基琥珀酰亚胺-α-萘乙酸酯(SINA)。研究了这一试剂在反相高效液相色谱中作为新的柱前衍生试剂,紫外可见检测,分离测定氨基酸的基本条件。衍生条件简单,方便。RP-HPLC分离了多种氨基酸,检测下限达pmol级。 展开更多
关键词 hplv 萘乙酸酯 氨基酸 SINA 柱前衍生试剂
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新疆啤酒花潜隐病毒的检测及外壳蛋白基因的序列分析 被引量:5
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作者 刘升学 韩盛 +2 位作者 祝建波 向本春 曹连莆 《植物病理学报》 CAS CSCD 北大核心 2008年第3期333-336,共4页
Hop samples from different region in Xinjiang were collected and detected by reverse transcription-polymerase chain reaction(RT-PCR) in order to determine the occurrence of HpLV.The RT-PCR products were 314 bp and 964... Hop samples from different region in Xinjiang were collected and detected by reverse transcription-polymerase chain reaction(RT-PCR) in order to determine the occurrence of HpLV.The RT-PCR products were 314 bp and 964 bp.Coat protein(CP) gene segments of isolates AW3 from Wensu,CH1 from Changji,K6 from Yanqi,SW8 from Shawan,TE2 from Emin,TY4 from Tacheng and YL4 from Yili were cloned.Nucleic acid sequence of the CP gene and deduced CP amino acid sequence alignments were compared with AB032469 registered in GenBank.The result showed that 8 HpLV isolates were divided into two groups.Among the isolates,AB032469,K6,AW3,CH1,TE2,TY4 and YL4 were classified as GroupⅠand SW8 was assigned to GroupⅡ.GroupⅠisolates fell into three small clusters.Nucleotide and deduced amino acid sequence comparisons within GroupⅠisolates showed that the sequence homologies were 98.1%-99.9% and 99.0%-100%,respectively. Comparisons of nucleotide and deduced amino acid sequences between Group Ⅰ and Group Ⅱ showed that the sequence homologies were 95.8%-96.4% and 98.4%-99.0%,respectively.Amino acid identity was higher than nucleic acid identity among all isolates. 展开更多
关键词 新疆啤酒花 潜隐病毒 外壳蛋白基因 序列分析 检测 VIRUS 寄主范围 酒花品种
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