Journal:

Article Title: TATA box-binding protein (TBP)-related factor 2 (TRF2), a third member of the TBP family

doi:

Figure Lengend Snippet: Comparison of TBP family members. (a) Percent identity in the bipartite repeat domain between the Drosophila TBP family members. (b) Organization of domains in TBP family members. The arrows represent the bipartite repeats of the core domain. (c) Alignment of the core domains of dTRF2, hTRF2, ceTRF2, and the TBP consensus sequence, with “X” denoting nonconserved residues. ∗ denotes residues conserved between TRF2 family members that are different from the TBP consensus. The TBP consensus was obtained by comparing Drosophila, human, and C. elegans TBP. Shown are residues 198–379 of dTRF2, 7–186 of hTRF2, and 178–466 of C. elegans TRF2. Full-length Drosophila TRF2 and full-lentgh human TRF2 have been deposited in the GenBank database (accession nos. AF136569 and AF136570). # in the ceTRF2 sequence denotes the 109-residue insertion.

Article Snippet: pGST-hTRF2 was constructed by inserting human TRF2 into pVL1392GST ( 14 ) using Nde I and Xba I sites generated by PCR. pGST-dTRF2 was constructed by inserting residues 149–375 of dTRF2 into pVL1392GST by using Nde I and Xba I sites generated by PCR. pGST-hTRF2 and pGST-dTRF2 were used for production of recombinant baculovirus (PharMingen) which was used to infect Sf9 cells and produce GST-hTRF2 and GST-dTRF2.

Techniques: Sequencing

Journal:

Article Title: TATA box-binding protein (TBP)-related factor 2 (TRF2), a third member of the TBP family

doi:

Figure Lengend Snippet: Biochemical analysis of TRF2. (a–d) Western blot analysis of TFIIA and TFIIB binding to TRF2. A GST fusion of dTRF2 (a and b) or hTRF2 (c and d) and GST (a–d) was bound to glutathione beads and then incubated with dTFIIA (a), dTFIIB (b), hTFIIA (c), or hTFIIB (d). Unbound protein was then washed away, and the bound protein was detected by Western blot analysis. (e) Elution of endogenous dTRF2 from a Superose 6 gel filtration column (Amersham Pharmacia). Endogenous Drosophila TRF2 eluted slightly after the 670-kDa molecular mass standard. The elution profile of the molecular mass standards is indicated along the axis.

Article Snippet: pGST-hTRF2 was constructed by inserting human TRF2 into pVL1392GST ( 14 ) using Nde I and Xba I sites generated by PCR. pGST-dTRF2 was constructed by inserting residues 149–375 of dTRF2 into pVL1392GST by using Nde I and Xba I sites generated by PCR. pGST-hTRF2 and pGST-dTRF2 were used for production of recombinant baculovirus (PharMingen) which was used to infect Sf9 cells and produce GST-hTRF2 and GST-dTRF2.

Techniques: Western Blot, Binding Assay, Incubation, Filtration

Journal:

Article Title: TATA box-binding protein (TBP)-related factor 2 (TRF2), a third member of the TBP family

doi:

Figure Lengend Snippet: Northern blot analysis of hTRF2 expression in multiple human tissues. (Upper) Blots were probed with TRF2 and actin. An alternative TRF2 transcript was observed in testis (∗), and an alternative actin transcript was observed in both muscle and heart (∗). (Lower) Comparison of relative hTRF2 levels. The hTRF2 signal was normalized against actin, and the testis level was arbitrarily set to 100. pbl, peripheral blood leukocyte.

Article Snippet: pGST-hTRF2 was constructed by inserting human TRF2 into pVL1392GST ( 14 ) using Nde I and Xba I sites generated by PCR. pGST-dTRF2 was constructed by inserting residues 149–375 of dTRF2 into pVL1392GST by using Nde I and Xba I sites generated by PCR. pGST-hTRF2 and pGST-dTRF2 were used for production of recombinant baculovirus (PharMingen) which was used to infect Sf9 cells and produce GST-hTRF2 and GST-dTRF2.

Techniques: Northern Blot, Expressing