Analysis of Genes for DNA Replication Enzymes and Homeobox Proteins in Congenital Multiple Anomalies.

1.DNA聚合酶Delta辅助蛋白和同源异源蛋白：我们分离了水稻增殖细胞核抗原的基因组克隆，并对其中一个克隆进行了部分测序。该序列与大鼠增殖细胞核抗原(P.N.A.S.86：3189-3193，)的氨基酸序列有60%的同源性。我们还测定了果蝇增殖细胞核抗原基因的全核苷酸序列，并在5^，-侧翼区发现了一簇可能与含有同源结构域的基因产物结合的位点。足迹分析表明，果蝇同源域蛋白(偶数跳过和零点)可以特异性地结合这些位点(M.C.B.10：872-879,1990)。为了证明人类发育程序性分化调控与细胞增殖调控之间的密切关系，我们分离了人增殖细胞核抗原和同源异型盒4A的基因组克隆。先天性畸形DNA复制酶基因和同源异型盒基因的研究：应用DNA印迹杂交技术，对15例先天畸形中的1例进行DNA印迹杂交，发现异常限制性片段。用DNA聚合酶β和γ-晶体作探针，在95例正常人中检出1例异常限制性片段。用Box4A和4B探针进行DNA印迹分析，在先天畸形的DNA样本中没有发现任何异常的限制性内切酶片段。对30个严重程度先天性畸形的DNA样本进行了伽马晶体DNA片段的多态性检测，该片段已被定位在Chomosome2的Homeobox 4基因座附近。在统计上，先天性畸形和伽马晶体Gan的RFLP之间存在显著的相关性。这一结果表明，与先天性畸形密切连锁的基因座存在于伽马晶体和同源盒4基因座(核酸符号)的附近。先生。1989年21：19-20，IRL出版社)。

1. DNA polymerase delta auxiliary protein (PCNA) and homeopox protein : We isolated genomic clones encoding rice PCNA and partially sequenced one of the clones. The rice amino acid sequence shared 60% homology with the sequence of the rat PCNA (P. N. A. S. 86 : 3189-3193, 1989). We also determined the c omplete nucleotide sequence of the Drosophila PCNA gene and found a cluster of putative binding sites for the homeodomain-containing gene products in the 5^,-flanking region. Footprint analysis revealed that Drosophila homeodomain proteins (even-skipped and zerknullt) can specifically bi these sites (M. C. B. 10 : 872-879, 1990). In order to demonstrate the close relationship between the regulation of developmentally programmed differentiation and that of cell proliferation in man, we isolated genomic clones for human PCNA and homeobox 4A.2. Studies on genes for DNA replication enzyme and on homeobox genes in congenital anomalies : Abnormal restriction fragments were detected by DNA blot hybridization with PCNA, DNA polymerase beta and DNA polymerase alpha probes in one of 15 cases of congenital malformations. Abnormal restriction frgments were detected in one case of 95 normal individuals when DNA polymerase beta and gamma-crystalline were used as probes. DNA blot analysis using box 4A and 4B probes did not show any unusual restriction fragments in the DNA samples of congenital anomalies. Thirty DNA samples of severe degree of congenital malformations were examined for polymorphisms with gamma-crystalline DNA fragment which has been mapped near the homeobox 4 locus in chomosome2. Statistically significant association was demonstrated between congenital malformations and RFLPs of gamma-crystalline ganes. This result indicates that the locus closely linked to congenital malformations is present in the proximity of gamma-crystalline and homeobox 4 loci (Nucleic Acids Symp. Ser. 21 : 19-20, 1989, IRL press).