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  3. Vol. 2 No. 1 (2016): Winter
  4. Research/Original Article- Immunology

ISSN: 2423-6241

Winter
Vol. 2 No. 1 (2016)

Cloning, expression and library construction for HIV-1 Tat Protein

  • Farzaneh Sheikholeslami
  • Niloufar Sadat Kalaki
  • Seyed Abdolhamid Angaji
  • Sahar Eshghjoo
  • Ali Reza Janani
  • Samereh Gholami Moghadam
  • Mehrdad Araiinejad
  • Kazem Baesi
  • Minoo Mohraz
  • Asghar Abdoli

Archives of Medical Laboratory Sciences, Vol. 2 No. 1 (2016),
https://doi.org/10.22037/amls.v2i1.13688 Published 31 October 2019

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Abstract

Background: Designing novel therapeutic agents has been a critical challenge for HIV disease.

Materials and Methods: In current study a DNA sequence which was encoded the Tat protein was synthesized and inserted in pET 28 vector. Vector was cloned in BL21-DE3 E. coli and cultured in TB media. After protein expression, recombinant Tat protein was purified by NTA affinity chromatography. The Tat purified protein efficiency and confirmed by SDS-PAGE and Western blot, respectively. We were immunized the camel against HIV-1 Tat recombinant protein to made a camelid antibody library. Total RNA was extracted from camel lymphocytes and VHH fragments synthesized and amplified using RT-PCR and Nested- PCR methods by special primers.

Results: The 350- 450 bp VHH gene fragment was produced by RT-PCR and Nested- PCR and extracted from agarose gel 1%. Then gel extraction was performed and pure fragments were inserted in HEN-4 vector by T4 DNA ligase.
Conclusion: The library can be applied for biopanning and isolation of nanobody against HIV-1 Tat Protein. Nanobody small size may be a useful drug for treatment of HIV disease because give them the potency of the recognizing the cryptic epitopes of tat and neutralized the virus.

Keywords:
  • HIV-1
  • Tat Protein
  • VHH
  • Nanobody
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How to Cite

1.
Sheikholeslami F, Kalaki NS, Angaji SA, Eshghjoo S, Janani AR, Gholami Moghadam S, Araiinejad M, Baesi K, Mohraz M, Abdoli A. Cloning, expression and library construction for HIV-1 Tat Protein. Arch Med Lab Sci [Internet]. 2019Oct.31 [cited 2023May28];2(1). Available from: https://journals.sbmu.ac.ir/archives/article/view/13688
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The journal of "Archives of Medical Laboratory Sciences" is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Print ISSN: ‪2423-6241; Online ISSN: 2476-7670

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