^(125)Ⅱ标记病毒DNA探针制备方法的研究  

Studies on the Methods of Preparation of ^(125)l-Labeling Viral DNA Probes

作　　者：谌济国 吴远明[2] 

机构地区：[1]江西医学院微生物教研室,330006 [2]中国科学院武汉病毒研究所,武汉430071

出　　处：《病毒学杂志》1990年第4期404-409,共6页

摘　　要：本文报道以油桐尺蠖核型多角体病毒(BsNPV)DNA、单纯疤疹病毒(HSV)DNA、λDNA为材料,用国产^(125)I-碘化钠制备探针。探讨了离子强度,pH值、反应温度及时间诸因素对以三氯化铊为氧化剂核酸标记的影响。在pH值为4.8—4.9,离子强度(以Na^+计)为0.1mol/L、反应温度为70℃、时间为30分钟条件下,可获得高比活性的产物。^(125)I的掺入率在10—30％之间,探针的比活性可超过10~6cpm/μgDNA。点杂交结果显示,探针最低可检测出1Pg同源DNA,探针的特异性强,存放于-20％冰箱的探针在两个月内最低检测量没有明显变化。By using BsNPV DNA, HSV DNA and λDNA, the paper reported the re suits of the influence of ionic strength, pH value, reaction temperature and time on the labeling process of DNA with radioactive iodine in the presence of thallic chloride.When pH value was 4.8—4.9, ionic strength(Na^+)was 0.1mol/L, the temperature was 70℃ and the reaction time was 30 minutes, very high specific activity can be yielded. The percentage of ^(12)Ⅰ bound to DNA is 10—30% and the specific activity of DNA probe is over 10~8 cpm/μg DNA. The DNA probes can detect as little as 1pg target DNA with high specificity. After stored in -20℃ for two months, the detecting ability of probes will not significantly change.

关 键 词：碘125 标记病毒 DNA探计 

分 类 号：Q93－332[生物学—微生物学]