PCP4 maps between D21S345 and P31P10SP6 on chromosome 21q22.2->q22.3
PCP4 maps between D21S345 and P31P10SP6 on chromosome 21q22.2->q22.3
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DOI：
10.1159/000134623
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发表时间：
1997-01-01
期刊：
影响因子：
--
通讯作者：
Korenberg, JR
中科院分区：
文献类型：
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作者：
Hubert, RS;Korenberg, JR
Materials and methodsTo assess potential roles of chromosome 21 genes in the Down syndrome phenotypes we have constructed contigs spanning D21S55 to MX 1 (Hubert et al., 1997) and isolated expressed sequences including thecDNA RH10B1 for PEP-19. The RH10B1 cDNA is 549 bp in length and shows high homolo gy to rat PEP-19 (GenBank accession number M24852) 81% over 499 bp. mouse PEP-19 (GenBank accession number X17320) 83% over 482 bp, human PEP-19 (GenBank accession number X93349) 98% over 502 bp. and to three ESTs (GenBank accession number HS090212. HS191160. HS164178). This cDNA was isolated using a direct cDNA selection proce dure (Yamakawa et al.. 1995) and the bacterial artificial chromosome (BAC) clone 423A5. BAC 423A5 maps to chromosome 2 Iq22. 2-> q22. 3 using fluo rescence in situ hybridization and just distal to D21S345 (see Fig. 1). RH 10B1was shown to map back to BAC 423A5 using Southern blot analysis of NotVEcoKI digested BAC 423A5 and RH 10B1insert as a probe (data not shown).To more precisely map the location of PCP4, PCR primers were designed specific to the 3'exon of the gene and PCR was performed on somatic cell hybrids, YACs, BACs and Pl-derived artificial chromosomes (PACs) spanning the region defined by 423A5. The primers: gac atl gac atg gat gca cc and act agg act gag ace cag cc amplify a 100-bp PCR product using the following mix and thermal cycling conditions. The 50 pi PCR reactions contained 2 pi of 1: 25 diluted clor. e mini-prep DNA; 0.9 pM each primer, 100 pM each dNTP, 1 x PCR buffer (Perkin Elmer. 10 mM Tris-HCl. 1.5 mM MgCh, 50 mM KCI pH 8.3). and 2.5 U AmpliTaq Gold polymerase (Perkin Elmer). PCR was performed in an MJ Research thermocvcler under the following conditions: DNA was initially denatured at 93 C for 9 min followed by 35 cycles of 93 C for I min. 60 C for 1min. and 72 C for 30 s. A final extension at 72 C was carried out for 3 min.
