Construction of a representative cDNA library from prostatic intraepithelial neoplasia.
Construction of a representative cDNA library from prostatic intraepithelial neoplasia.
复制标题
前列腺上皮内瘤变代表性 cDNA 文库的构建。
作者：
D. Krizman;Rodrigo F Chuaqw;P. Meltzer;J. Trent;P. Duray;W. Linehan;L. Liotta;M. Emmert;E
我们报道了从前列腺上皮内瘤变中分离的显微切割细胞构建的基于质粒的cDNA库。提取总RNA，通过寡核苷酸(DT)介导的逆转录和连接子的添加转化为钝端的双链cDNA.用带有UDG相容末端的连接子特异的引物扩增该基因，并将扩增产物亚克隆。对154个克隆进行了测序，结果表明，81.5%的克隆来自已知基因、匿名表达序列标签或新转录本，而筛选克隆的冗余度很小。这些结果证明了从代表癌症进展的微观前驱阶段的特定显微解剖细胞群中构建复杂的有代表性的cDNA文库的可行性。这种方法应该有助于鉴定在不同组织来源和肿瘤形成阶段的细胞中特异表达的转录本。
We report the construction of a plasmid-based cDNA library made from microdissected cells derived from prostatic intraepithelial neoplasia. Total RNA was extracted and converted to blunt-ended, double-stranded cDNA by oligo(dT)-mediated reverse transcription followed by linker addition. A linker-specific primer with UDG-compatible ends was used to amplify the cDNA and the resulting PCR product was subcloned. A total of 154 clones were sequenced and results indicated that 81.5% of the clones derived from either known genes, anonymous expressed sequence tags, or novel transcripts with very little redundancy of screened clones. These results demonstrate the feasibility of constructing complex representative cDNA libraries from specific microdissected cell populations that represent microscopic precursor stages of cancer progression. This method should facilitate identification of transcripts specifically expressed in cells of a distinct histological origin and tumorigenic stage.
LNCaP 前列腺癌细胞系中的 t(6;16)(p21;q22) 染色体易位导致 tpc/hpr 融合基因。
DOI：
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发表时间：
1996
期刊：
Cancer research.
影响因子：
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作者：
Veronese,ML;Bullrich,F;Negrini,M;Croce,CM
通讯作者：
Croce,CM
影响因子：
56.9
作者：
DONG, JT;LAMB, PW;BARRETT, JC
通讯作者：
BARRETT, JC