Dooley T P, Obermoeller R D, Leiter E H, Chapman H D, Falany C N, Deng Z, Siciliano M J
Department of Genetics, Southwest Foundation for Biomedical Research, San Antonio, Texas 78228.
Genomics. 1993 Nov;18(2):440-3. doi: 10.1006/geno.1993.1494.
We have recently cloned a cDNA encoding the human phenol-preferring phenol sulfotransferase (P-PST) enzyme. An oligonucleotide primer pair based on the human STP (representing sulfotransferase, phenol-preferring) cDNA sequence was synthesized and was employed in polymerase chain reaction (PCR) amplification of human genomic DNA to identify a 525-bp DNA fragment. The DNA sequence of this portion of the STP gene, near the 5' end of the coding region, was determined. The amplified genomic fragment contained two small introns of 104 and 89 bp. When DNA samples from a human-hamster somatic cell hybrid panel were screened by PCR using these primers, only those hybrids that contained human chromosome 16 were positive for the 525-bp genomic fragment. To identify the specific region on chromosome 16 that contained the STP gene, PCR amplification reactions were performed on a human-mouse somatic cell hybrid panel containing defined portions of human chromosome 16. The results indicated that STP is localized proximal to the gene for protein kinase C, beta 1 polypeptide (PRKCB1), in the region from the distal portion of 16p11.2 to p12.1. The human STP gene maps near the locus for Batten disease (CLN3). Furthermore, we have determined by genotyping of murine interspecific backcross progeny that the homologous gene in mouse (Stp) localizes to the syntenic region of mouse chromosome 7 near the D7Mit8 (at 54 cM) and D7Bir1 markers.
我们最近克隆了一个编码人偏爱苯酚的酚磺基转移酶(P-PST)的cDNA。基于人STP(代表磺基转移酶,偏爱苯酚)cDNA序列合成了一对寡核苷酸引物,并将其用于人基因组DNA的聚合酶链反应(PCR)扩增,以鉴定一个525bp的DNA片段。测定了STP基因编码区5'端附近这部分的DNA序列。扩增的基因组片段包含两个分别为104bp和89bp的小内含子。当使用这些引物通过PCR筛选人-仓鼠体细胞杂交板的DNA样本时,只有那些包含人16号染色体的杂交体对525bp的基因组片段呈阳性。为了鉴定16号染色体上包含STP基因的特定区域,对包含人16号染色体特定部分的人-小鼠体细胞杂交板进行了PCR扩增反应。结果表明,STP定位于16p11.2远端至p12.1区域中蛋白激酶Cβ1多肽(PRKCB1)基因的近端。人STP基因定位于贝敦氏病(CLN3)基因座附近。此外,我们通过对小鼠种间回交后代进行基因分型确定,小鼠中的同源基因(Stp)定位于小鼠7号染色体的同线区域,靠近D7Mit8(54cM)和D7Bir1标记。