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Journals / Gülhane Tıp Dergisi / 2009 / Cilt: 51 - Sayı: 2

Expression of HBeAg gene region in Saccharomyces cerevisiae

HBeAg gen bölgesinin Saccharomyces cerevisiae'da ekspresyonu

Mehmet YAPAR
Kenan ŞENER
ORHAN BEDİR
ERTAN ALTAYLI
Ayhan KUBAR
Pages
91–93
DOI
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

HbeAg producing yeast cells obtained may render it possible to purely isolate proteins in future studies.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:HBeAg/anti-HBe seroconversion is one of the markers frequently used in the diagnosis of hepatitis B infection.
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Key Findings & Data:In this study, the “e” antigen gene region of the hepatitis B virus (HBV) was transformed to Saccharomyces cerevisiae and expressed by the yeast in order to obtain HBeAg protein used in serologic tests.
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Conclusion & Impact:HbeAg producing yeast cells obtained may render it possible to purely isolate proteins in future studies.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

HBeAg/anti-HBe seroconversion is one of the markers frequently used in the diagnosis of hepatitis B infection. In this study, the “e” antigen gene region of the hepatitis B virus (HBV) was transformed to Saccharomyces cerevisiae and expressed by the yeast in order to obtain HBeAg protein used in serologic tests. Firstly, HBeAg gene region was amplified by PCR method. Later, the amplicons were cloned into the plasmid pYES2.1 via the TOPO® TA expression kit and transformed into the competent bacteria (TOPO10F' Escherichia coli). The plasmid pYES2.1+HBeAg was isolated from the competent bacteria, which was cultivated on LB media supplemented with ampicillin, and transformed to Saccharomyces cerevisiae via the S.c. Easy- Comp Transformation Kit. Finally, whether HBeAg was expressed by the yeast was checked through two different automatized systems. HbeAg producing yeast cells obtained may render it possible to purely isolate proteins in future studies.

Özet

HBeAg/anti-HBe serokonversiyonu hepatit B enfeksiyonlarının tanısında sık kullanılan göstergelerden birisidir. Bu çalışmada serolojik testlerde kullanılan HBeAg proteinini elde etmek amacıyla hepatit B virüsü (HBV) “e” antijen gen bölgesi Saccharomyces cerevisiae'ya aktarıldı ve eksprese ettirildi. Bunun için öncelikle HBeAg gen bölgesi PCR yöntemiyle çoğaltıldı. Daha sonra amplikonlar TOPO® TA ekspresyon kiti ile pYES2.1 plazmidine klonlandı ve kompetan bakteriye (TOPO 10F' Escherichia coli) transforme edildi. Ampisilinli LB agarda çoğaltılan kompetan bakteriden pYES2.1+HBeAg plazmidi izole edildi ve S.c. EasyComp transformasyon kitiyle Saccharomyces cerevisiae'ya transforme edildi. Son olarak, HBeAg'nin maya tarafından eksprese edilip edilmediği iki farklı otomatize sistem aracılığıyla kontrol edildi. Elde edilen HBeAg üreten maya hücreleri sonraki çalışmalarda üretilen proteinlerin pür olarak elde edilmesine imkan sağlayabilir.

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Cite This Article

YAPAR, M., ŞENER, K., BEDİR, O., ALTAYLI, E., KUBAR, A. (2009). Expression of HBeAg gene region in Saccharomyces cerevisiae. Gülhane Tıp Dergisi, Cilt: 51 - Sayı: 2, 91–93. https://ajindex.com/articles/225478

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References (11)

  1. Badur S. Hepatit A, B ve D virusları. In: Ustaçelebi Ş, Abacıoğlu H, Badur S (eds). Moleküler, Klinik ve Tanısal Viroloji. Ankara: Güneş Kitabevi, 2004: 175-202
  2. Harrison TJ, Dusheiko GM, Zuckerman AJ. Hepatitis viruses. In: Zuckerman AJ, Banatvala JE, Schoub BD, Griffiths PD, Mortimer P (eds). Principles and Practice of Clinical Virology. 6th ed. London: John Wiley & Sons Ltd, 2009: 273-319
  3. Seeger C, Zoulim F, Mason WS. Hepadnaviruses. In: Knipe DM, Howley PM (eds). Fields Virology. 5th ed. New York: Lippincott Williams & Wilkins, 2007: 1253-1304
  4. Yapar M, Aydogan H, Pahsa A, et al. Rapid and quantitative detection of Crimean-Congo Hemorragic Fever Virus by one step real-time reverse transcriptase PCR. Jpn J Infect Dis 2005; 58: 358-362
  5. Yapar M, Guney C, Basustaoglu AC, Kubar A. Expression of hepatitis B “e” antigen gene in Escherichia coli. Mikrobiyol Bul 2001; 35: 273-278. 6. http://structure.biochem.queensu.ca/protocols/ compcells.htm (Son erişim tarihi: 17.03.2008) 7. pYES2.1 TOPO® TA Expression Kit. Five-minute cloning of Taq polymerase-amplified PCR products for regulated expression in Saccharomyces cerevisiae. Catalog no. K4150-01. Invitrogen. User manual
  6. Guthrie CC, Fink GR. Guide to yeast genetics and molecular biology. In: Abelson JN, Simon MI (eds). Methods in Enzymology. Vol
  7. San Diego: Academic Press, 1991
  8. Saracli MA, Sener K, Gonlum A, Yildiran ST, Wickes BL. Genotyping of clinical Rhodotorula mucilaginosa isolates by pulsed field gel electrophoresis. Mycoses 2003; 46: 487-491
  9. Kniskern PJ, Hagopian A, Montogomery DL, et al. Unusually high level expression of foreign gene (hepatitis B virus core antigen in Saccharomyces cerevisiae). Gene 1986; 46: 135-141
  10. Kubar A, Yapar M, Ozyurt M, Haznedaroglu T, Gun H. Cloning of hepatitis B virus surface gene region to Escherichia coli. Flora 1988; 3: 183-186
  11. Yapar M, Guney C, Saracli MA, Kilic A, Kubar A. Expression of HBsAg gene in insect (Trichiplusia ni) cell culture. Flora 2001; 6: 108-113.