Cell Lines

BHK cells(clone 13)(Macpherson and Stoker1962) and Hela cells were kindly provided by the Imperial Cancer Research Fund, London

ND7 cells(Wood et al, 1990) were kind gifts from Dr. J. Wood, Sandoz institute, London

DRG cells were isolated from adult Wistar strain rats.

II. 1.4. Tissues

Rat tissues were obtained from 10-week-old Wistar strain rats for RNA extraction.

II.1.5. DNA (plasmids)

1)Bluescript vector SK^(Stratagene)

2) Bluescript kS plasmid containing the full length Oct-2.5 cDNA was provided by Dr. S. Dawson.

3) Bluescript KS plasmids contain the coding sequences for the mutants o f the Oct-2 repressor domain(ME, MEL, MKR, MRKL, MEKR, MERE AND MERQ).

4) pGEX-2TK vector containing the GST gene(Stratagene).

5) pGEX-0ct2RD plasmid containing a cDNA fragment encoding the Oct-2 repressor domain(amino acids 142-181 of Oct-2) linked to the GST gene via EcoR I cloning site(provided by Dr. K. Lillycrop).

6) Eukaryotic expression vectors:

G5E1BCAT contains five Gal4 binding sites and the ElB TATA box driving the expression of the CAT gene(Carey et al., 1990).

03G5E1BCAT was made by cloning three copies of the overlapping octamer/T AAT GARAT motif( AT GCT A AT GARAT) upstream of the Gal4 binding sites in the vector G5E1BCAT.

pSG424 vector contains the SV40 ori/early promoter region fused to the coding sequence for Gal4(l-147), followed immediately by a polylinker and translation stop codons(Sadowski and Ptashne 1989).

pSG424 vector containing the C-terminal acidic activation domain of VP 16 fused in frame to the Gal4 DNA-binding domain was provided by Professor M. Ptashne(Carey et al., 1990).

pSG424 vector containing the full length coding sequence of Gal4 was provided by Dr. U.M. Hansen(Licht et al,, 1993)

pSG424 vector containing the full length cDNA of E l A fused in frame to the Gal4 DNA binding domain was provided by Dr. M.R. Green(Lillie and Green 1989).

pSG424 vector containing the C-terminal acidic activation domain of NF^B linked to the Gal4 DNA-binding domain was provided by Dr. P. Baeuerle(Schmitz et al., 1994).

pSG424 vector containing the C-terminal activation domain of SPl linked to the Gal4 DNA-binding domain was provided by Dr. U.M. Hansen(Licht et al., 1993).

pSG424 vector containing the N-terminal activation domain of c-Fos linked to the Gal4 DNA-binding domain was provided by Dr. T. Kouzarides(Brovm et al., 1995).

pSG424 vector containing the full length cDNA of TAT linked to the Gal4 DNA-binding domain was provided by Dr. L. Lania(Southgate and Green 1991).

The construct containing the SV40 ori/early promoter region fused to the coding sequence for Gal4(l-93) was provided by Professor W. Schaffher(Seipel et al., 1992).

activation domain of CTF linked to the Gal4 DNA-binding domain(Gal4 1-93) were provided by Professor W. Schaffner(Seipel et al., 1992).

The constructs containing the N-terminal or C-terminal domain of Oct-2 linked to the Gal4 DNA-binding domain(Gal4 1-93) were provided by Professor P. Matthias(Friedl and Matthias 1995)

pRPXG4 vector is modified from the expression vector pSG424. The original EcoRl site has been destroyed and another site introduced in frame with the EcoRl site in the mouse glucocorticoid receptor at amino acid 506.

pRPXG4 vectors containing the coding sequences for the Oct-2 repressor domain(amino acids 141-202) or the Oct-1 amino acids 224-272 linked to the Gal4 DNA-binding domain were provided by Dr. K. Lillycrop(Lillycrop et al., 1994a).

pRPXG4 vectors contain the coding sequences for the mutants of the Oct-2 repressor domain(ME, MEL, MKR, MRKL, MEKR, MERE AND MERQ) linked to the Gal4 DNA-binding domain via EcoR 1 cloning site.

pJ7 expression vector(Morgenstem and Land, 1990) contains the constitutive immediate- early cytomegalovirus(CMV) IE72 promoter flanked 3' by a polylinker, an intron and a transcriptional termination signal.

pJ7 vector containing the isolated POU domain of Oct-2 was provided by Dr. K. Lillycrop(Lillycrop et al., 1994a).

The construct containing the constitutive immediate-early cytomegalovirus(CMV)IE94 promoter driving the expression of Oct-2.5 was provided by Dr. T. Wirth(Wirth et al.,

1991).

The constructs containing the constitutive immediate-early cytomegalovirus(CMV)IE94 promoter driving the expression of full length Oct-2.2 and various deletions of Oct-2.2 were provided by Professor P. Matthias(Muller-Immergluck et al., 1990).

The construct containing the constitutive immediate-early cytomegalovirus(CMV)IE94 promoter driving the expression of truncated Oct-2 (154-376 amino acids) was provided by Dr. T. Gerster(Gerster et al., 1990).

pBLCAT2 contains a multiple cloning site flanking the HSV thymidine kinase(TK) promoter from-105 to +51 driving the expression of the CAT gene.

pBLCAT2 plasmids containing various fragments of the HSV IE-3 promoter upstream of the HSV TK promoter were cloned by Dr. S. Dawson(Dawson et al., 1996b)

pBLCAT2 containing the Oct-2 binding site AT GCTAAT G AG AT upstream of the HSV TK promoter was provided by Dr. K. Lillycrop(Morris et al., 1994).

pBL2CAT expression vectors containing five Gal4 binding sites cloned at position -105, -770 or +1000 relative to the transcriptional start site of HSV TK promoter were provided by Dr. P.M. Rauscher (Madden et al., 1993).

The reporter constructs designated T7G5-TATA, T7G5-TATA-Inr and T7G5-Inr contain the CAT gene under the control of adenovirus major late promoter(AdMLp) Inr and/or the ElB TATA box with five Gal4 DNA-binding and seven tetO sequences(provided by Dr. L. Lania)(Pengue and Lania 1996).

ptetR containing the C-terminus of prokaryotic protein tetR encoded by TnlO from Escherichia coli was provided by Dr. L. Lania(Pengue and Lania 1996).

ptetR expression vector contains the coding sequences for the Oct-2 repressor

domain(amino acids 141-202) linked to the tetR DNA-binding domain via EcoR 1 cloning site.

ptetR vectors contain the coding sequences for the mutants of the Oct-2 repressor domain(MEL, MKR, MRKL, MEKR, MERE AND MERQ) linked to the tetR DNA- binding domain via EcoR 1 cloning site.

The reporter constructs designated G5SV, G5SV and G51 lOSV contain the CAT gene under the control of the simian virus 40(SV40) enhancer-less promoter with the 5 Gal4 DNA-binding sites inserted in both orientation immediately upstream or 110 bp upstream the three 21 bp repeats of SV40 early promoter(provided by Dr. L. Lania)(Pengue et al., 1994).

The reporter constructs designated G5-38HIV, G5HIV, G5LTRHIV and LTRHIVG5 contain the CAT gene under the control of the HIV-1 promoter with the 5 Gal4 DNA- binding sites inserted at-38, -83, at -770 from the HIV-1 start site and at the 3' of the CAT gene(provided by Dr. L. Lania)(Pengue et al., 1994).

The TAT expression plasmid(pSV-TAT) contains the full length cDNA o f TAT under the control of the SV40 early region promoter(provided by Dr.L. Lania)(Pengue et al., 1994).

The TAT expression plasmids contain the TAT cDNAs mutated at cysteine 37(TAT37), cysteines 22 and 37(TAT22/37) and with the N-terminal 48 amino acids deleted under the control of the SV40 early region promoter(provided by Dr.L. Lania)(Pengue et al., 1995).

The reporter plasmid LTRHIVCAT contains a CAT gene under the control of the full length HIV-1 LTR(provided by Dr. L. Lania)(Pengue et al., 1995).

The pCG expression vector contains the human CMV |JE promoter, the HSV TK gene 5' untranslated leader and initiation codon, rabbit p-globin gene splicing and polyadenylation

signals and the replication origin of SV40.(Tanaka and Herr 1990).

The pCG expression vectors containing the full length cDNAs of Oct-1 and Oct-2 were provided by Dr. W. Herr(Tanaka and Herr 1990).

The pCG expression vetcors containing the Oct chimaeras were also kind gifts from Dr. W. Herr. These constructs were prepared by combining Oct-1, Oct-2, and their deletion derivatives at the engineered Xho I and/or Sal I sites(Tanaka and Herr, 1990).

IE-1 CAT plasmid contains the HSV IE-1 promoter(ffom -585 to +150) driving expression of the CAT gene(provided by Dr. S. Solverstein)(Gelman and Silverstein

1987).

IE-3 CAT plasmid contains the HSV IE3 promoter(from -330 to +33) driving expression of the CAT gene(provided by Dr. C. Preston)(Stow et ah, 1986).

PLTR poly(American Type Culture Collection, U.S.A.) contains the mononey murine leukaemia virus(MoMuLV) promoter which has been modified by deletion of a cryptic splice site in the SV40 3' untranslated region (Gorman, 1985).

PLTR poly expression vectors containing the full length cDNAs of Bm-3a(long and short isoforms), Bm-3b(short) were provided by Dr. T. Theil(Theil et al, 1993).

PLTR poly expression vector containing the Bm-3 chimaeras were also kind gifts from Dr. T. Theil. These constructs were prepared by the ligation of different segments of Bm- 3a and Bm-3b sequences generated by PCR(Theil et al., 1993, Theil and Moroy, 1994).