Structural and functional analysis of the human Y-box binding protein (YB-1) gene promoter
Structural and functional analysis of the human Y-box binding protein (YB-1) gene promoter
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DOI：
10.1093/nar/24.10.1873
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发表时间：
1996-05-15
影响因子：
14.9
通讯作者：
Kohno, K
中科院分区：
文献类型：
--
作者：
Makino, Y;Ohga, T;Kohno, K
我们已经分离了三个重叠的基因组克隆，其中含有人YB-1基因的5'部分。这些克隆跨越类似于25 kb的连续DNA，含有10 kb的5'侧翼序列和15 kb的基因。测定第一外显子和上游2000个碱基对（bp）的核苷酸序列。第一个外显子异常大，包含166 bp的编码序列和331 bp的非翻译区。YB-1基因5 '端的CpG序列包括第一个外显子和内含子以及上游区域，第一个外显子附近的GC含量约为70%，非翻译序列中有一个无CpG区。在主要转录起始位点之前的片段不包含TATA盒、CCAAT盒和已知转录因子的结合序列。使用氯霉素乙酰转移酶（CAT）基因的瞬时表达测定表明，从+24至+281的序列对于CAT表达是关键的。荧光原位杂交证实YB-1基因位于染色体1 p34。对其它噬菌体基因组DNA的聚合酶链反应分析表明，有几个克隆来自假基因。
We have isolated three overlapping genomic clones containing the 5' portion of the human YB-1 gene. These clones span similar to 25 kb of contiguous DNA containing 10 kb of 5' flanking sequence and 15 kb of the gene. The nucleotide sequence of the first exon and of 2000 upstream base pairs (bp) was determined. The first exon is unusually large and contains a 166 bp coding sequence and a 331 bp untranslated region. CpG sequences cover the 5'-end of the YB-1 gene including its first exon and intron as well as the upstream regions, The GC content around the first exon is similar to 70% and a CpG-free region was located in the untranslated sequence. The segment preceding the major transcription initiation site does not contain a TATA box, CCAAT box and the binding sequence for known transcription factors. A transient expression assay using the chloramphenicol acetyltransferase (CAT) gene showed that the sequence from +24 to +281 was critical for CAT expression. Fluorescence in situ hybridization demonstrated the chromosomal locus of YB-1 gene on chromosome 1p34. Polymerase chain reaction analysis on other genomic phage DNAs showed that several clones were derived from pseudogenes.