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本文设计并化学合成了一对PCR引物,以人CD4cDNA为模板,成功地扩增出人CD4N端两个结构域的编码基因(600bp),并在此基因两端分别插入了EcoRI和HindⅢ酶切位点及起始和终止码。通过EcoRI和HindⅢ双酶切位点将CD4基因克隆入pUC19质粒,经过PCR和酶切鉴定得到CD4重组克隆pT403。DNA序列分析结果表明,所克隆CD4基因与已发表的人CD4基因序列完全一致。将CD4基因 相似文献
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The human immunodeficiency virus type 1 gag p20 gene fragment was clonedinto plasmid pBV220 to construct a recombinant expression plasmid pCY7.By induction,the p20 protein was expressed in E.coli,and it was confirmed by Western blotting thatthe expressed p20 protein could be specifically recognized by anti-p17 monoclonalantibodies.The recombinant bacteria was lysed and fractionated into snpernatant andprecipitate by centrifugation.It was found that the p20 protein was present chiefly inthe precipitate.After p20 protein being washed twice,its purity reached 27.4%.Then theprecipitate was washed with 1 mol/L urea solution and the purity of p20 protein wasraised to 58.1%.Finally,the precipitate was dissolved in 6mol/L of urea and appliedonto a heparin affinity column.Four peaks were obtained and the p20 protein wasfound mainly in peak 3.The purity of p20 protein at this step reached 84.2% as meas-ured by gel scanning. 相似文献
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Burkitt淋巴瘤伴有t(8;14),t(8;8)或t(8;22)易位,90%的滤泡淋巴瘤伴有t(14;18)易位。这些易位均涉及到免疫球蛋白重链或轻链基因位点和MYC或bc1-2和bc1-1基因的重组,重组MYCbc1-2或bc1-1基因的高水平表达是B细胞瘤发生的重要原因。 相似文献
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作者应用位于探针DNA两端的两个DNA片段作引物,以探针DNA为模板,在大肠杆菌DNA聚合酶I的介导下,经过1—3次变性、退火和延伸过程,使探针DNA得到标记.3次实验结果表明,应用此法标记的DNA探针的比活性可以达到2×10~(14)cpm/g DNA,标记效率大于60%;而用缺刻翻译法标记的DNA探针,其标记效率只有22.7%.表明这是一种较好的DNA探针标记方法. 相似文献
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经抗AFP iRNA预处理的小鼠脾细胞,在体外培养的第一天即出现AFP特异性抗体生成细胞,iRNA在125~250μg/ml间活性最高,此活性能被RNase破坏而不受DNase和pronase影响。免疫荧光试验表明,抗AFP iRNA能诱导小鼠脾细胞表达膜AFP抗体或受体。 相似文献
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