乙型肝炎表面小抗原(sHBsAg)的克隆与表达

Christian Heryakusuma, Fernita Puspasari, Ihsanawati Ihsanawati, E. Giri-Rachman, M. Tan, Eka Ramadhani, N. Nurainy, D. Natalia
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引用次数: 2

摘要

重组乙型肝炎表面小抗原(sHBsAg)被用作预防乙型肝炎病毒感染的疫苗成分。短句来源为构建局部重组sHBsAg,采用pcr扩增的印度尼西亚sHBsAg B基因型和adw2亚型DNA片段,通过重组方法将其克隆到Hansenula polymorpha表达载体pHIPX4中。pHIPX4-sHBsAg被整合到多态H. NCYC495基因组的乙醇氧化酶(AOX)位点,并在多态H. AOX启动子的控制下调控sHBsAg的表达。携带sHBsAg编码序列的H. polymorpha NCYC495在无机物培养基中培养,加入0.5% (v/v)的甲醇诱导重组sHBsAg的表达。采用HBsAg诊断试剂盒、ELISA和Western blot检测sHBsAg的表达。
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Cloning and Expression of Small Hepatitis B Surface Antigen (sHBsAg) In Hansenula polymorpha
Recombinant small hepatitis B surface antigen (sHBsAg) is used as a vaccine component to prevent hepatitis B virus infection. As an attempt to produce local recombinant sHBsAg, a PCR-amplified DNA fragment encoding Indonesia sHBsAg which belongs to B genotype and adw2 subtype was cloned into Hansenula polymorpha expression vector pHIPX4 by using recombination method. The resulted pHIPX4-sHBsAg was integrated into the alcohol oxidase (AOX) locus of H. polymorpha NCYC495 genome and the sHBsAg expression was regulated under the control of H. polymorpha AOX promoter. H. polymorpha NCYC495 carrying the sHBsAg coding sequence was grown in mineral medium and methanol 0.5% (v/v) was added to induce the expression of recombinant sHBsAg. The expression of sHBsAg was detected by HBsAg diagnostic kit test, ELISA, and Western blot analysis.
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