Application of an improved cDNA competition technique to identify prostate cancer-associated gene.
Application of an improved cDNA competition technique to identify prostate cancer-associated gene.
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应用改进的 cDNA 竞争技术鉴定前列腺癌相关基因。
DOI：
10.1089/104454999314827
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Steiner,MS
中科院分区：
文献类型：
--
作者：
Rinaldy,AR;Steiner,MS
利用来自高转移MAT-LyLu和低转移AT-1 Dunning R3227大鼠两个密切相关的cDNA群体的混合探针，开发了一种改进cDNA文库筛选的技术 前列腺癌亚系。该技术需要从每个亚系产生cDNA文库，然后通过聚合酶链反应（PCR）扩增cDNA插入群体。pcr产物 将来自第一文库的PCR产物进行放射性标记，并与过量的来自第二文库的PCR产物混合。使用该混合物和过量的λ和pBluescriptDNA 作为探针筛选第一个cDNA文库。该混合探针（命名为竞争探针）与筛选的第一cDNA文库的噬菌斑剥离物差异性地交叉杂交。弱放射性 信号表明竞争探针混合物和第一cDNA文库共有的cDNA序列的交叉杂交，而强信号则意味着未杂交的独特或丰富的cDNA序列 在第一个cDNA文库中。通过显示全长cDNA克隆与筛选的第一细胞系的表型相关来证实该技术的再现性。的分离的 克隆的特征为大鼠核仁蛋白、编码16 S和12 S rRNA的大鼠线粒体基因以及对缬氨酸和苯丙氨酸特异的大鼠tRNA。这一结果与以下事实相一致： 第一种细胞系MAT-LyLu在代谢上比AT-1细胞更有活性，因为更高的基因剂量或核仁和线粒体RNA及其相关基因的扩增。另一个克隆 其具有强信号，代表与MAT-LyLu癌症表型相关的新基因。
A technique to improve cDNA library screening was developed by using mixed probes derived from two closely related cDNA populations of high-metastatic MAT-LyLu and low-metastatic AT-1 Dunning R3227 rat prostate cancer sublines. The technique required the generation of a cDNA library from each subline followed by polymerase chain reaction (PCR) amplification of the cDNA insert population. The PCR products derived from the first library were radiolabeled and mixed with an excess amount of PCR products from the second library. The mixture and an excess amount of both the lambda and pBluescript DNA were used as a probe to screen the first cDNA library. This mixed probe (designated the competition probe) differentially cross-hybridized with the plaque lift of the screened first cDNA library. Weak radioactive signals indicated the cross-hybridization of cDNA sequences common to the competition probe mixture and the first cDNA library, whereas strong signals implied unhybridized unique or abundant cDNA sequences in the first cDNA library. The reproducibility of this technique was confirmed by showing that the full-length cDNA clones were associated with the phenotype of the screened first cell line. The isolated clones were characterized as rat nucleolar protein, rat mitochondrial genes coding for 16S and 12S rRNAs, and rat tRNAs specific for valine and phenylalanine. This result is consistent with the fact that the first cell line, MAT-LyLu, is metabolically more active than are AT-1 cells because of higher gene dosage or amplification of nucleolar and mitochondrial RNA and its associated genes. Another clone which had a strong signal represented a novel gene associated with the MAT-LyLu cancer phenotype.
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色素性干皮病互补 A 组相关基因：使用抗环丁烷二聚体和 (6-4) 光产物的单克隆抗体进行确认。
DOI：
10.1016/0921-8777(93)90065-o
发表时间：
1993
期刊：
Mutation research
影响因子：
--
作者：
Mori,T;Rinaldy,TL;Athwal,RS;Kaur,GP;Nikaido,O;Lloyd,RS;Rinaldy,A
通讯作者：
Rinaldy,A
影响因子：
64.5
作者：
MATHER, EL;ALT, FW;KOSHLAND, ME
通讯作者：
KOSHLAND, ME
影响因子：
14.9
作者：
LIANG, P;AVERBOUKH, L;PARDEE, AB
通讯作者：
PARDEE, AB
在差异竞争杂交策略中使用 cDNA 文库进行基因克隆：应用于克隆 XP-A 相关基因。
DOI：
10.1089/dna.1.1988.7.563
发表时间：
1988
期刊：
DNA (Mary Ann Liebert, Inc.)
影响因子：
--
作者：
Rinaldy,A;Dodson,ML;Darling,TL;Lloyd,RS
通讯作者：
Lloyd,RS
通过杂交竞争对克隆的发育调控盘基网柄菌基因进行选择和分析。
影响因子：
14.9
作者：
G. Mangiarotti;S. Chung;C. Zuker;H. Lodish
通讯作者：
H. Lodish