用于检测肿瘤病毒特异性DNA序列的RNA-DNA滤膜杂交法的可靠性。
Reliability of the RNA-DNA filter hybridization for the detection of oncornavirus-specific DNA sequences.
作者信息
Shoyab M, Markham P D, Baluda M A
出版信息
J Virol. 1974 Aug;14(2):225-30. doi: 10.1128/JVI.14.2.225-230.1974.
Abstract
Denatured DNA from leukemic myeloblasts or uninfected chicken embryos, immobilized on nitrocellulose filters, was hybridized to a vast excess of [(3)H]70S RNA from purified avian myeloblastosis virus. The viral RNA was eluted from the RNA-DNA hybrids, purified, and then rehybridized in solution to an excess of either leukemic or normal chicken embryonic DNA. This study revealed that all the slow and the fast hybridizing viral RNA sequences detectable by liquid hybridization in DNA excess had hybridized to the filter bound DNA. Both techniques also gave similar values for the number of 28S ribosomal RNA genes contained in a chicken cell genome: 210 by the liquid hybridization procedure and 218 by the filter hybridization technique. Therefore, filter hybridization can accurately detect DNA sequences present in relatively few numbers in the genome of higher organisms.
摘要
固定在硝酸纤维素滤膜上的来自白血病成髓细胞或未感染鸡胚的变性DNA，与大量过量的来自纯化禽成髓细胞瘤病毒的[(3)H]70S RNA进行杂交。病毒RNA从RNA-DNA杂交体中洗脱、纯化，然后在溶液中与过量的白血病或正常鸡胚DNA再次杂交。这项研究表明，在DNA过量的情况下通过液相杂交可检测到的所有慢速和快速杂交病毒RNA序列都已与滤膜结合的DNA杂交。两种技术对于鸡细胞基因组中所含28S核糖体RNA基因的数量也给出了相似的值：液相杂交法为210个，滤膜杂交技术为218个。因此，滤膜杂交能够准确检测高等生物基因组中数量相对较少的DNA序列。
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