Affinity labelling and cloning of steroid and thyroid hormone receptors / edited by H. Gronemeyer.

Date:
[1988]
Catalogue details

Licence: Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)

Credit: Affinity labelling and cloning of steroid and thyroid hormone receptors / edited by H. Gronemeyer. Source: Wellcome Collection.

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    Ch. 16] Analysis of the human glucocorticoid receptor gene promoter 301 Ch*r«cl«ri9ation of human GR gene promoter Smal GRE PstI I I Ì ^ Ï % тт - rao-oi Y • âM-au-ot u-&•> Anil mti Hr *UctlNt •МЙ ' SS7- О ъ S С а о. М/ •ззв' 3-CCGCCOCCCTTOACGCCT-S Ollgonucitolld« uttd for icrtaning •Ad prtmtr txltntton -661 -651 -641 -631 -621 -611 -601 -591 -581 CCCAATGTGC TTTACACTTT ACTAGACATC CGGCTCGATC АСТОТТСТДТ GCAACTTGCT GCAGACGGAT AACCAATTTC TCAGAGCACA GRE -571 -561 -551 -541 -531 -521 -511 -501 -491 TAGCTATCGA CGCGATTGAT GTGAGCGCAC ACTTCCGGCA GGACGTGGCT ATGGGGGGCG AAGTGCGCAT CTGACGAGGT CCGGCACCCT -481 -471 -461 -451 -441 -431 -421 -411 -401 GCAGCCTCGG CCGGTCGGGC CCCCGAGGCT GGGGGCAGAC CCGGCGCCCC TGCAGTTGCC AACGTCACCA ACAGTGCATC GTTCCXCGCC -405 -391 -381 -371 -361 -351 -341 -331 -321 -311 gîCGCGGCCC TCGGGCGGGG AGCGGCCGGG GGTGGAGTGG AGCGCGTGTG TGCGAGTGTG TGCCCGTGCG CCGCCTCCAC GCCCTCCCCT -388 -301 -291 -281 -271 -261 -251 -241 -231 -221 CGGTCCCGCT CGCTCCGAGG CCGGGCTGCC CTTCGCTGTC CGCGCTCTCT TCCCTCCGCC GTCGCCGCTC CATTTTGCGA GCTCTTCTGT -249 -211 -201 -191 -181 -171 -161 -151 -141 -131 GACGGACCCA GTCACGCCTC TCGGGATGGA TTCTTGGTGG AAGGAGACCC GCACCGGAGC GGCGCGAAGC AGCTGGGACC GGGACGGCAC -121 -111 -101 -91 -81 -71 -61 -51 -41 GCGCGCCCGG AAGCCCCGAC CCGCGGAGCC GGCCGCGGCG GAGGGCTGGC TTGTCAGCTG GGCAATGGGA GACTTTCTTA AATAGGCTCT -95 -31 -21 -11 -1 +11 +21 +31 +41 4: +51 CCCCCCACCC ATGGAGAAAG gGGCGGCTGT TTACTTCCTT TTTTTAGAAA AAAAAAATAT ATTTCCCTCC TGCTCCTTCT GCGTTCACAA Ncol -20 I ^ * Î Transcription 1 елоп— +91 +101 +111 ^ +121 +131 +141 GCTAAOTTGT TTATCTCGGT CGCGGCGGGA ACTGCŒTATAGCTAATCGAA GAGTATTGCT GACAACGGTT CAACTTTCCG TGATGGCACA j/GCGCCGCCCT ТСАСсфт./ * primer 338 ^-íntron +151 +161 +171 +181 +191 +201 +211 GACTCTTTTA CTCGTTGGCC TCCACTGACC TATAAAAACA CTTCTGCAGG ATTCTGGCGT ACCGTTCCTG TCTAGA. Xbtl Fig. 2 — Top, partial structure of the human glucocorticoid receptor gene. Top left, a restriction map of the promoter region showing the start site of transcription, located in the Smal-Smal fragment which has been introduced into the Bluescribe vector (described on the right), and the open reading frame starting in exon III. Bottom, nucleotide sequence of the 5' portion of HGR gene where the arrow corresponds to the transcription initiation site. D. Nuclease SI mapping The Smal-Smal (1.6 kb) genomic subfragment (Fig. 2) inserted into the BSM13 (BS(SmaI-SmaI 1.6)) at the 3' to 5' orientation to the T7-RNA polymerase and linearized with Ncol (31 bp 5' to the transcription initiation; see Fig, 2) was used in synthesizing a uniformly [^^P]CTP labeled anti-mRNA probe (see Fig. 2). When this