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Objective To analyze the effect of the identification and evaluation of Escherichia (E.) coli and Shigella,based on the upstream flanking sequences of CRISPR1.Methods Both CRISPR and cas sequences were obtained through the BLAST with repeating sequences against the publicly complete genome in GenBank that related to E.coli and Shigella.Clustal X was used to perform multi-sequences alignment of the flanking sequences.PCR method was used to amplify the upstream flanking sequences of CRISPR1 in order to appraise the effect of identification and evaluation of upstream flanking sequences on E.coli and Shigella,which were based on the upstream flanking sequences of CRISPR1.Results The results showed that 73.4％ of the strains containing the I-E CRISPR/Cas that belonged to the phylogroups A,B1,D while 8.4％ strains carried the I-F CRISPR/Cas.Another 17.2％ of the strains owned CRISPR3-4 (non-CRISPR/Cas) only belonged to the phylogroups B2.All the Shigella strains carried I-E CRISPR/Cas.More than 99％ of similarity the CRISPR1 upstream-flanking sequences was seen in E.coli (except B2) and Shigella and E.coli (B2).Both sensitivity and specificity were greater than 91％ after PCR amplification in the region to identify the E.coli and Shigella.Conclusion The upstream of CRISPR1 could achieve a preliminary identification effect on E.coli and Shigella.
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Objective To analyze the effect of the identification and evaluation of Escherichia (E.) coli and Shigella,based on the upstream flanking sequences of CRISPR1.Methods Both CRISPR and cas sequences were obtained through the BLAST with repeating sequences against the publicly complete genome in GenBank that related to E.coli and Shigella.Clustal X was used to perform multi-sequences alignment of the flanking sequences.PCR method was used to amplify the upstream flanking sequences of CRISPR1 in order to appraise the effect of identification and evaluation of upstream flanking sequences on E.coli and Shigella,which were based on the upstream flanking sequences of CRISPR1.Results The results showed that 73.4％ of the strains containing the I-E CRISPR/Cas that belonged to the phylogroups A,B1,D while 8.4％ strains carried the I-F CRISPR/Cas.Another 17.2％ of the strains owned CRISPR3-4 (non-CRISPR/Cas) only belonged to the phylogroups B2.All the Shigella strains carried I-E CRISPR/Cas.More than 99％ of similarity the CRISPR1 upstream-flanking sequences was seen in E.coli (except B2) and Shigella and E.coli (B2).Both sensitivity and specificity were greater than 91％ after PCR amplification in the region to identify the E.coli and Shigella.Conclusion The upstream of CRISPR1 could achieve a preliminary identification effect on E.coli and Shigella.
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Objective To analyze the sequence of microsatellite and the flanking sequence from four populations of Oncomelania hupensis. Methods We cloned 159 SSR-PCR amplification products of a commonly used primer, (CA)8RY, using O. hupensis genomie DNA as template, and sequenced 82 products Results The sequences obtained were novel O. hupensis genomic sequences but not repeat simple sequence. It was observed that 36 out of 82 clones contained microsatellites between priming sites.The flanking sequences of certain microsatellite were invariant. Both (GA/CT). and (TTAGGG/CCCAA)n were found in four populations of O. hupensis. However, (CAA)n were found only in O. hupensis from Fuqing,Fujian province and (TCTCTG), were found only in O. hupensis from Guichi,Anhui province and (GAA/TTC)n, (CAA/TTG)n, (CAT), were found only in O.hupensis from Puge,Sichuan province. Conclusion The results obtained by SSR-PCR should not be interpreted as the amplification of microsatellite loci, and analytical rules similar to those for Random Amplified Polymorphic DNA should be used. SSR-PCR could not make the most of the priority of microsatellite. It seems better to amplify the microsatellites with the primers designed on the basis of the flanking sequence.
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Abstract Objective:To better understand the regulatory mechanism of Pax-5 gene expression on B-cell differentation and development.Methods:The isolation and genomic colning of the 5'-flanking region of Pax-5 gene were carried out according to established protocols: molecu-lar cloning and subcloning The nucleotide sequences were determined by a double-stranded dideoxy sequencing method. Sequence analysis wasperformed on line programs. Results:Analysis of tha total sequence(6 671 bp) shows the proximal promoter include 3 CAT boxes, 1SP1 and 1Ebox. However, there was no consensus sequence for a TATA box in the 5'-flanking region. Putative regulatory sites of further upstream in thesequence revealed 6LMO2 COM, 5NFAT, 2LPOLYA-B, 3GATA1, 2AP4, 10MZF1, 1ets1B, 1GATA3, 1NKX-25, 2RORA1, 1LYF1, 2Ikaros2,2TCF11 , 1GATA-C and 1FREAC7. Conclusion: The 5'-flanking regionn of human Pax-5 gene exon1B could be involved in regulating the expres-sion of human Pax5 and B-cell differentiation and development.