应用光敏生物素探针检测杂交瘤及细胞中的小鼠白血病病毒

Detection of MuLv in Hybridoma and other cell line with Photobiotin-labeled Probe

采用ＢａｍＨＩ酶切和琼脂糖凝胶电泳技术，从重组质粒ＰｓＦｌｌ中回收５ｋｂ小鼠白血病病毒（ＭｕＬＶ）ＤＮＡ片段，经光敏生物素标记后制备ＤＮＡ探针，以斑点杂交法检测杂交瘤细胞、ＭｃＡｂ纯品以及ＳＰ２／０细胞中的ＭｕＬｖ。结果表明：此探针灵敏度可达２５ｐｇ，特异性好，在被检测的１３株杂交瘤细胞和２株ＳＰ２／０细胞中分别查出５株和１株为ＭｕＬｖ阳性，ＭｃＡｂ纯品中未发现ＭｕＬｖ。

MuLV DNA fragment(5kb)in recombinant plasmid,Which wasdigested with restriction enzyme,was isolated by agarose gel electrophoresis and labeled with photobiotin as a probe. The probe was used to detect MuLv in hybridoma cells,purified McAb products and SP2/C cells with dot hyoridization.The result showed that the probe issensitive and specific to detect MuLv under 25 pg homologous DNA. In total of l3 hybridoma and 2 SP2/O cells detected.the positive rate was 38%and 50%respectively. There was no MuLv DNA founded inp uri- fiedMcAb products.