• Title/Summary/Keyword: precore/core mutants

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Role of pre-C Region in the Expression and Secretion of Hepatitis B Viral Core Antigen in Yeast (효모에서 B형 간염바이러스의 내면항원의 발현과 분비에 미치는 전위내면항원의 역할)

  • 신상훈;김성기;노현모
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.1-5
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    • 1990
  • The coding sequence of hepatitis B viral core antigen (HBcAg) (subtype adr) contains two in-phase initiation codons, one for precore and the other for core antigen gene. To study the expression of core antigen and the role of precore region, the coding sequence of HBcAg with or without precore (pre-C) region were subcloned into yeast expression vector containing phosphoglycerate kinase (PGK) promoter. To study the role of upstream region in the expression of the core antigen, a series of 5' deletion mutants were also subcloned into the vector. After transformation into various host strains, the expression of HBcAg were analysed by radio-immunoassat. Under optimal condition of core antigen gene expression in yeast, the highest amount of antigen was detected in the cell line SHY4 containing pGKHBc plasmid composed of the yeast PGK gene promoter, terminator and C-gene. Regardless of the presence of precore region, core antigen was not detected in the medium but in cell extract. These results suggest that precore region cannot affect the secretion of core antigen in Saccharomyces cerevisiae.

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Low Frequency of Precore Mutants in Anti-Hepatitis B e Antigen Positive Subjects with Chronic Hepatitis B Virus Infection in Chennai, Southern India

  • Shanmugam, Saravanan;Velu, Vijayakumar;Nandakumar, Subhadra;Madhavan, Vidya;Shanmugasundaram, Uma;Shankar, Esaki Muthu;Murugavel, Kailapuri G.;Balakrishnan, Pachamuthu;Kumarasamy, Nagalingeswaran;Solomon, Suniti;Thyagarajan, Sadras Panchatcharam
    • Journal of Microbiology and Biotechnology
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    • v.18 no.10
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    • pp.1722-1728
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    • 2008
  • The natural course of chronic hepatitis B (CH-B) virus infection is reportedly variable, and the long-term outcomes in hepatitis B e antigen (HBeAg)-negative chronic hepatitis B infection are distinct from HBeAg-positive chronic hepatitis. However, the molecular virological factors that contribute to the progression of liver disease in the south Indian setting remain largely unclear. We prospectively studied 679 consecutive patients for HBsAg, HBeAg, anti-HBe, and HBV DNA by qualitative PCR. Randomly selected samples were subjected to bidirectional sequencing to reveal core/precore variants. Of the total 679 chronic HBV cases investigated, 23% (154/679) were replicative HBV carriers. Furthermore, amongst the 560 HBV DNA samples analyzed, 26% (146/560) were viremic. Among the 154 HBeAg positive cases, HBV DNA was positive in 118 cases (77%), significantly (p<0.001) higher than the anti-HBe positive (7%) (28/406) cases. Significant increase in liver disease (p<0.01) with ALT enzyme elevation (p<0.001) was observed in both HBe and anti-HBe viremic cases. Interestingly, low frequencies of mutations were seen in the precore region of the HBV strains studied. HBV precore and core promoter variants were less often detected in subjects with "e" negative chronic HBV infection and, therefore, may not have a prognostic role in determining liver disease sequelae in this part of tropical India.