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为筛选脑发育相关的基因 ,应用新发展的抑制消减杂交技术 ,以新生大鼠小脑mRNA逆转录cDNA作为测试者 (tester) ,成年大鼠小脑mRNA逆转录cDNA作为驱动者 (driver) ,进行消减杂交 ,克隆制备成新生大鼠小脑高表达的cDNA文库。挑选出 5 0个重组质粒 ,测序得到 37个不同基因片段序列。再次用反Northern杂交确认有意义的差异表达基因。同时将测序结果与GenBank注册序列进行同源性比较 ,2 0个大鼠中首次测得的基因表达序列标签 (EST)已被GenBank收录 (BG6 95 72 6 -BG6 95 74 5 )。
In order to screen the genes involved in brain development, the newly developed suppression subtractive hybridization technique was used. The reverse transcription cDNA of cerebellum mRNA of neonatal rats was used as a tester, and the reverse transcription cDNA of adult rat cerebellar mRNA was used as a driver for subtraction Hybridization, cloning prepared into neonatal rat cerebellum highly expressed cDNA library. Fifty recombinant plasmids were selected and sequenced to obtain 37 different gene fragment sequences. Northern blotting was again used to confirm meaningful differentially expressed genes. At the same time, the sequencing results were compared with the GenBank sequence. Genome-based ESTs (GenBank accession numbers) were obtained from 20 of the 20 rats (BG6 95 72 6 -BG6 95 74 5).