促髓细胞分化的锌指基因单向PCR扩增直接测序的研究

锌指基因是一种造血调节基因，编码锌指结构蛋白，主要在髓细胞中表达，促进髓细胞分化，在急性早幼粒白血病维甲酸治疗中，促使病情缓解。本文报道了我们从基因分子上研究锌指基因作用中，探索并建立了单向聚合酶链反应（ＰＣＲ）扩增特定单链ＤＮＡ，直接测序的新方法。它能产生质和量均佳的单链ＤＮＡ，无需纯化即可直接用于测序，使复杂的测序研究简便易行，可在２，３天内完成。这种单向ＰＣＲ扩增特定单链ＤＮＡ直接测序的方法，经对锌指基因的ｃＤＮＡ测序，得到验证。此法不仅适用于疾病研究中的ＤＮＡ测序，还可制各单链ＤＮＡ探针，更利于基因结构组成的研究。

Generation of Single Stranded DNA by Unidirectional PCR and Its Application to Direct Sequencing of The Human Myeloid Zinc Finger Gene (MZF-1)

The human myeloid zinc finger gene 1(mzf-1) is a hematopoitic regulatory gene that encodes a zinc finger protein, expressed preferentially in myeloid cells, and is essential for granulocytic differentiation. We report here an unidirectional PCR method for the generation of singlestranded DNA in studies of mzf-1. This unidirectional amplification method produces high yield of virtually pure ssDNA products. Since the quantity and quality of the ssDNA are very high, the ssDNA products can be used directly in DNA sequencing without any major purification. Using this method of ssDNA production, DNA sequence can be obtained within 2 to 3 days. The application of the unidirectional amplification method is demonstrated by the analysis of mzf-1 cDNA.This efficient method is suitable for DNA sequence analysis, for the preparation of ssDNA probes, and potentially useful in studies of genomic structural organization.