Schematic representation of the PCR DNA fragments ThyA-F, ThyA-Δ3′, ΔThyA and X-thyAΔ3′-X′

<p><b>Copyright information:</b></p><p>Taken from "Efficient and seamless DNA recombineering using a thymidylate synthase A selection system in "</p><p>Nucleic Acids Research 2005;33(6):e59-e59.</p><p>Published online 30 Mar 2005</p><p>PMCID:PMC1072810.</p><p>© The Author 2005. Published by Oxford University Press. All rights reserved</p> () PCR amplification using primers thyA-F1 and thyA-R1 () amplifies a 1470 bp fragment (thyA-FL) of the gene containing the promoter, Factor Sigma 54 binding site and the transcription termination flanking the start and stop codons for translation. The 1124 bp product, thyA-Δ3′, amplified using primers thyA-F1 and thyA-R3 (), lacks 347 bp 3′ of the transcription terminator. ΔthyA is a 744 bp product generated by overlapping PCR (see Materials and Methods for details) with most of the coding sequence deleted (+123 to +848; relative to the translational start site), and Δ in the diagram represents the deleted region. X-thyAΔ3′-X′ is a fusion product amplified using primers ColX-thyAF and ColX-thyAR with 5′ and 3′ overhang sequences for the regions of the gene for homologous recombination. () Schematic representation of the strategy for the generation of the ColX-G18D fragment. Two PCR products were first generated using primer sets, ColX-F/ColX-mR and ColX-mF/ColX-R. These products, containing complementary sequences from primers ColX-mF and ColX-mR, were used in an overlapping PCR to produce ColX-G18G using primers ColX-F and ColX-R. () Schematic representation for the amplification of a PCR product from a plasmid, pG18D-Flag, using primers ColX-F and ColX-R. pG18D-Flag contains mutations (bold and underlined) around the signal peptide cleavage site of collagen X, and a Flag sequence inserted at the 3′ region of exon 2 (shaded region)

Relationship between rs430397 G>A distribution and liver functional profiles in patients with liver cirrhosis.

<p>SD, standard deviation; T-Bil, total bilirubin; ALB, albumin; ALT, alanine aminotransferase; AST, aspartate aminotransferase; GGT, γ-glutamyltransferase; PT, prothrombin time.</p><p>*Compared with the GG group, <i>x²</i> = 4.461 and <i>P</i> = 0.029.</p>†<p>Compared with the GG group, <i>x²</i> = 5.832 and <i>P</i> = 0.016.</p

Comparison between advanced and mild liver cirrhosis patients according to Child-Pugh score.

<p>*Mann-Whitney U-test.</p>†<p>Pearson Chi-square test.</p>‡<p>Multivariable logistic regression analysis adjusted with age and gender.</p>§<p>Child-Pugh classification.</p

Genotype distribution of rs430397 G>A in the fifth intron of the <i>GRP78</i> gene in HBV-infected individuals with different HBeAg states.

<p>CHB, chronic hepatitis B; CHC, chronic hepatitis B virus carrier; HBeAg, hepatitis B e antigen; HBV, hepatitis B virus; IHC, inactive hepatitis B surface antigen carrier; LC, liver cirrhosis.</p><p>*Compared with the phase I group, <i>x²</i> = 7.562 and <i>P</i> = 0.006; and compared with the phase III group, <i>x²</i> = 5.969 and <i>P</i> = 0.015.</p>†<p>Compared with the phase I group, <i>x²</i> = 8.039 and <i>P</i> = 0.005; and compared with the phase III group, <i>x²</i> = 4.883 and <i>P</i> = 0.027.</p>‡<p>Compared with the phase I group, <i>x²</i> = 7.131 and <i>P</i> = 0.008.</p

Demographic characteristics and haplotypic distributions of the 3′ untranslated region (UTR) of <i>GRP78</i> gene among cases with HCC.

<p>*Chi-square test.</p><p>AFP, <i>α</i>-fetoprotein; HBV, hepatitis B virus; HCC, hepatocellular carcinoma; TNM, tumor, nodes, metastasis-classification.</p

rs430397 G>A polymorphism and cirrhosis risk in HBV patients.

<p>*Pearson Chi-square test.</p>†<p>Multivariable logistic regression analysis adjusted with age and gender.</p

Clinical and laboratory features of the subjects included in the study.

<p>ALB, albumin; ALT, alanine aminotransferase; AST, aspartate aminotransferase; CHB, chronic hepatitis B; CHC, chronic hepatitis B virus carrier; CT, computed tomography; GGT, γ-glutamyltransferase; HBV, hepatitis B virus; IHC, inactive hepatitis B surface antigen carrier; LC, liver cirrhosis; MRI, Magnetic resonance imaging; PT, prothrombin time; SD, standard deviation;. T-Bil, total bilirubin.</p

Kaplan-Meier survival curves show that overall survivals of 576 cases with HCC were not associated with the different alleles, genotypes, haplotypes and diplotypes in the <i>GRP78</i> gene including rs16927997, rs1140763 and rs12009.

<p><i>P</i> value was calculated using a log-rank test. The wildtype alleles, homozygotes, and the corresponding haplotype and diplotype were designated as the referent.</p

Univariate survival analyses of possible prognostic factors.

<p>AFP, <i>α</i>-fetoprotein; HBV, hepatitis B virus; NS, not significant; TNM, tumor, nodes, metastasis-classification.</p