Gene expression analysis from human immunodeficiency virus type 1 subtype C promoter and construction of bicistronic reporter vectors

Nagadenahalli Byrareddy Siddappa, Venkatesh Prasanna Kashi, Mohanram Venkatramanan, Anangi Balasiddaiah, Narayana Jayasuryan, Anita Mahadevan, Anita Desai, Kadappa S. Satish, Susarla K. Shankar, Vasanthapuram Ravi, Udaykumar Ranga

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

We report the cloning and sequence analysis of the long terminal repeat (LTR) of several primary HIV-1 subtype C strains of India. Phylogenetically, all the LTRs and the paired env sequences clustered with subtype C reference strains. The LTRs demonstrated extensive polymorphism in the transcription factor binding sites (TFBS) within the enhancer and the modulator regions. We generated reporter vectors under the control of a select subset of the subtype C LTRs. The reporter vectors are distinguished by the simultaneous expression of two independent reporter genes, secreted alkaline phosphatase (SEAP) and enhanced green fluorescence protein (EGFP), in response to Tat. Expression of EGFP was facilitated by engineering an internal ribosome entry site (IRES) into the expression cassette. Although subtype C strains cause a large majority of the global infections, and important differences in the transcription factor binding sites have been identified in the subtype C promoter, few reporter vectors containing subtype C-LTR have been described. We analyzed gene expression from the C-LTR reporter vectors in different cell lines under diverse experimental conditions and compared it to the B-LTR reporter vector. The reporter vectors were responsive to Tat derived from diverse viral subtypes. Furthermore, a positive correlation was observed between the expression of the reporter genes and the viral structural protein p24 when the cells were infected with viral molecular clones. The LTR reporters we developed could be of significant use in the study of viral transactivation, in the evaluation of biological properties of viral subtypes, and in the screening for antiviral inhibitors.

Original languageEnglish (US)
Pages (from-to)1268-1278
Number of pages11
JournalAIDS Research and Human Retroviruses
Volume23
Issue number10
DOIs
StatePublished - Oct 1 2007

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Terminal Repeat Sequences
HIV-1
Gene Expression
Reporter Genes
Transcription Factors
Fluorescence
Binding Sites
Viral Structural Proteins
Transcriptional Activation
Antiviral Agents
Alkaline Phosphatase
Sequence Analysis
Organism Cloning
India
Proteins
Clone Cells
Cell Line
Infection

ASJC Scopus subject areas

  • Immunology
  • Virology
  • Infectious Diseases

Cite this

Gene expression analysis from human immunodeficiency virus type 1 subtype C promoter and construction of bicistronic reporter vectors. / Siddappa, Nagadenahalli Byrareddy; Kashi, Venkatesh Prasanna; Venkatramanan, Mohanram; Balasiddaiah, Anangi; Jayasuryan, Narayana; Mahadevan, Anita; Desai, Anita; Satish, Kadappa S.; Shankar, Susarla K.; Ravi, Vasanthapuram; Ranga, Udaykumar.

In: AIDS Research and Human Retroviruses, Vol. 23, No. 10, 01.10.2007, p. 1268-1278.

Research output: Contribution to journalArticle

Siddappa, NB, Kashi, VP, Venkatramanan, M, Balasiddaiah, A, Jayasuryan, N, Mahadevan, A, Desai, A, Satish, KS, Shankar, SK, Ravi, V & Ranga, U 2007, 'Gene expression analysis from human immunodeficiency virus type 1 subtype C promoter and construction of bicistronic reporter vectors', AIDS Research and Human Retroviruses, vol. 23, no. 10, pp. 1268-1278. https://doi.org/10.1089/aid.2006.0305
Siddappa, Nagadenahalli Byrareddy ; Kashi, Venkatesh Prasanna ; Venkatramanan, Mohanram ; Balasiddaiah, Anangi ; Jayasuryan, Narayana ; Mahadevan, Anita ; Desai, Anita ; Satish, Kadappa S. ; Shankar, Susarla K. ; Ravi, Vasanthapuram ; Ranga, Udaykumar. / Gene expression analysis from human immunodeficiency virus type 1 subtype C promoter and construction of bicistronic reporter vectors. In: AIDS Research and Human Retroviruses. 2007 ; Vol. 23, No. 10. pp. 1268-1278.
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AU - Balasiddaiah, Anangi

AU - Jayasuryan, Narayana

AU - Mahadevan, Anita

AU - Desai, Anita

AU - Satish, Kadappa S.

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AU - Ravi, Vasanthapuram

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