HIV-1 Reverse Transcriptase, Functional
05-001200 units,
05-0021000 units
Shipping and Storage: Ship at 4℃ or-20℃ and store at -20℃. Do not freezebelow -20℃
Product: Recombinant full-size HIV-1 Reverse Transcriptase without Tag-peptide expressedin E. coli. It is composed of heterodeimer, p66 and p51 as the one produced in HIV-1
infected cell. It is an RNA-dependent DNA polymerase derived from HIV-1 (AIDS virus),
subtype B origin (Ref.1). It also has RNaseH activity and is an enzyme indispensable forreproduction of AIDS virus.
Applications1)

It is extremely effective for screening new specific inhibitors for HIV virus as a drug fortreating AIDS).

2)

Generally, Gag and Env proteins are employed as antigens for detecting anti-HIV-1
antibody. However, by using this enzyme in combination as an antigen, the detection
will be more sensitive.

3)Standards for SDS-PAGE (Fig.1), Western blotting (Fig.2), Dot blotting, ELISA

Definition of activity: Activity of intake of 1 nmole of dTMP in 10 min at 37℃ is consideredas 1 unit using poly(rA) and oligo(dT) as template and primer.
Conditions of measurement: 50 mM Tris-HCl (pH 8.3), 10 mM MgCl2, 50mM KCl, 3 mM
DTT, 0.1% Nonidet P-40, 20 ug/ml poly(rA) ・ oligo(dT)12-18, 0.5 mM dTTP([3H]dTTP ,~1 x 105 cpm), and 10-50 units/ml reverse transcriptase.
Activity: ~5,000 units/ml
Purity: Over 90% by SDS-PAGE (CBB staining)
Form: 0.5 mg/ml in 50% glycerol, 40 mM Tris-HCl (pH8.3), 50 mM NaCl, 5 mM MgCl2, 0.1%
Triton X-100, 1 mM DTT
Data Link ：UniProtKB: P04585 GenBank: AAA44988.1

Fig. SDS-Polyacrylamide gel electrophoresisof HIV-1 reverse transcriptase

BioAcademia,Inc.

Tel. 81-6-6877-2335

Fax. 81-6-6877-2336

info@bioacademia.co.jp https://www.bioacademia.co.jp/en/

Fig.2. Western blotting of functional recombinant full-lengthHIV-1 reverse transcriptase by using anti-HIV-1 Reverse
Transriptase antibody (BioAcademia 65-001).
1; 40 ng / lane
2; 20 ng / lane3;

4 ng / lane4;

2 ng /lane

Anti-HIV-1 RT antibody was used at 1/2,000 dilution. As second
antibody, goat anti-rabbit IgG antibody conjugated with HRPwas used at 1/5,000 dilution. ECL system was used.

References: This product was described in Ref 1 and used in the following publications.
1. Saitoh A et al. Overproduction of human immunodeficiency virus type I reverse
transcriptase in Escherichia coli and purification of the enzyme. Microbiol Immunol 34:
509-521 (1990) PMID: 1699113
2.Permanasari ED et al. Enzymatic Activities of RNase H Domains of HIV-1 Reverse
Transcriptase with Substrate Binding Domains of Bacterial RNases H1 and H2. Mol
Biotechnol. 2015 Jun;57(6):526-38. PMID: 25673083
3. Kadokura K et al. Novel urushiols with human immunodeficiency virus type 1 reverse
transcriptase inhibitory activity from the leaves of Rhus verniciflua. J Nat Med. 2015
Jan;69(1):148-53. PMID: 25349048
4.Tada K et al. Abacavir, an anti-HIV-1 drug, targets TDP1-deficient adult T cellleukemia. Sci Adv. 2015 Apr 24;1(3):e1400203. PMID: 26601161
5.Izumida M et al. The Spirocyclic Imine from a Marine Benthic Dinoflagellate,Portimine, Is a Potent Anti-Human Immunodeficiency Virus Type 1 Therapeutic Lead
Compound Mar Drugs. 2019 Aug 24;17(9). PMID: 31450557
Useful Referece: Ref 1 describes infectious cDNA of HIV-1 which was used to constructexpression system of this product.
1. Adachi A et al ” Production of acquired immunodeficiency syndrome-associatedretrovirus in human and nonhuman cells transfected with an infectious molecularclone.” J Virol 59: 284-291 (1986) PMID: 3016298

BioAcademia,Inc.

Tel. 81-6-6877-2335

Fax. 81-6-6877-2336

info@bioacademia.co.jp https://www.bioacademia.co.jp/en/