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Jang, Jeong Won,Chun, Ji‐,Yong,Park, Young Min,Shin, Soo‐,Kyung,Yoo, Wangdon,Kim, Soo‐,Ok,Hong, Sun Pyo Blackwell Publishing Ltd 2012 Cancer Science Vol.103 No.2
<P>This study explored the combined effect of number and pattern of mutations in the X/precore regions of the hepatitis B virus (HBV) genome, mutational complex genotype (MCG), on hepatocellular carcinoma (HCC) development. Sequence variations were determined by direct sequencing and multiplex restriction fragment mass polymorphism analysis in 150 age‐, sex‐ and hepatitis B e antigen (HBeAg) status‐matched patients with and without HCC. In addition, a longitudinal study and an external validation of MCG were conducted. All were HBV subgenotype C2. Eight high‐frequency mutations (G1613A, C1653T, T1753V, A1762T, G1764A, A1846T, G1896A and G1899A) were significantly associated with HCC. Whereas C1653T, T1753V, G1764A and A1846T were independent mutational factors for HCC, the significance of these individual mutations was negligible when analyzed with all clinico‐virological variables. The total number of mutations was the only independent viral factor for HCC, irrespective of HBeAg status. There was a significant dose–risk relationship between the number of mutations and HCC, in which high risks for HCC were associated with mutation numbers ≥6. Pattern analysis of the mutations revealed disparity in distribution among the top seven high‐risk mutation combination patterns, which accounted for 40 and 2.7% of HCC and non‐HCC cases, respectively. The predictive accuracy of the high‐risk mutations for HCC was similar to that of α‐fetoprotein. Longitudinal and external validation studies also supported the association of mutation number with HCC development. MCG in the HBV X/precore regions is a risk indicator for HCC, and might serve as a new guide to the HCC screening scheme for chronic HBV carriers. (<I>Cancer Sci</I> 2012; 103: 296–304)</P>
Kim, Nam Keun,Hwang, Seong Gyu,Hong, Sun Pyo,Rhee, Hwanseok,Chung, Hyun Jae,Kim, Sukjoon,Han, Jin Hee,Kim, Hyung Tae,Rim, Kyu Sung,Kang, Myung Seo,Yoo, Wangdon,Kim, Soo-Ok 한국유전학회 2003 Genes & Genomics Vol.25 No.1
Lamivudine has been shown to be an effective therapy for chronic hepatitis B, but resistance to this agent is common during prolonged treatment. The predominant lamivudine resistance mutations are amino acid substitution in tyrosine, methionine, aspartate, aspartate (YMDD) motif of DNA polymerase including M552Ⅰ and M552Ⅴ. Current methods of detecting the HBV mutations are time-consuming, labor intensive, and not suitable for screening large numbers of samples. In the present study, we described the development of a genotyping assay able to screen viral variants and to identify and characterize and unidentified variation in a sensitive, cost-effective, and robust high-throughput manner. The assay is based on polymerase chain reaction (PCR) amplification and mass measurement of the oligonucleotides containing variation sites of the viral polymerase gene using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). When assessed for low limit of detection and for the ability to distinguish wild-type and variant DNA in mixed populations, this genotyping assay is sensitive to detect extremely low amounts of viral variants that correspond to 100 copies of HBV genome per ml and more sensitive than DNA sequencing in determining the ratios of wild-type to variant virus, as few as 1% of mutant among wild-type virus. When sera from 20 patients, each containing identified HBV mutants were analyzed, new MALDI-TOF MS-based assay exactly identified each variant and wild-type virus, and could detect additional viral genotypes that were not detected by conventional restriction fragment length polymorphisms (RFLP) method. In conclusion, the level of sensitivity and the amenability to high-throughput system along with intrinsic ability to identify an exact haplotype composed of two or more variations in a strand of DNA should lend the MALDI-TOF MS-based assay to the mass screening of HBV patients receiving lamivudine for further understanding virological response to therapy and disease progression.
형질전환체 담배에서 사람 파필로마 바이러스 HPV - 18 E6 유전자의 발현
전재필,현미아,김달웅,진승원,박순희,김현수,유왕돈,손우익 ( Jaepil Jeon,Meeah Hyun,Dalung Kim,Seungwon Jin,Soonhee Park,Hyunsoo Kim,Wangdon Yoo,Uik Sohn ) 생화학분자생물학회 1994 BMB Reports Vol.27 No.5
To develop a HPV-18 oral vaccine, HPV-18 E6 gene was cloned into an expression vector, under the control of CaMV 35S dual promoter and 5`UT region of TEV. The constructed vector (pUS-E6) was used to transform tobacco plants by the Agrobacterium-mediated leaf disc method. The transformants were then regenerated to whole transgenic plants. The HPV-18 E6 gene was found to be expressed in the transgenic tobacco plants as determined by PCR, RT-PCR, Northern analysis and Western analysis. These results show a possibility that HPV oral vaccine may be developed by expressing the HPV proteins in tomato or other vegetables.