Use of reporter gene in promoter analysis of the even-skipped (eve) gene

Restriction map of eve genomic DNA Smal Ell Pst‘l 1.65 1.5 0.05 Mlul Sacl NdelAsullEcoRV Kpnl BamHl Xhol HinclIXhol Hll XI Stil Tr n r’ t' nst rt 't kb 7.3 6.5 6.3 5.9 5.45 4.65 3.8 2.9 2.6 1.75 1.15 0.60.3 a ‘C'P'° a "9 IacZ coding Aseriesoftransoipfionalflisions wflhalarz “29'0" reporter gene are created using restriction enzymes . to remove pans of the regulatory sequence. and Deletion constructs assayed for expression in stripes 2 and 3. . . Fusron Expressron In construct stripe 2 stripe 3 5’A w + S’F 5’G S’H 5’l AB AC AD AE AF AG Al A1 + I++ I++l++++l¥++ l+++++l+l l Deletion analysis localizes enhancer modules to specific DNA sequences. ‘ ' 2"9 {‘7 1:0 Transcription start site Enhancer modules of eve genomic DNA 318 Stripe 3 Stripe 2 I led coding r l i . : region I [ To test deletion analysis results. a u’ansaiptional l l fusion of the stripe 2 enhancer module is l assayedwrththelacheportergene Stripe 2 [ad coding enhancer region module drives expression only in stripe 2. 1 Blue color is derived from the l activity of the lad reporter gene, l Brown color is derived from a 1 process called immunolocalizaaon l using an antibody specific to the ‘ eve protein. Stripezl 2 3 4 5 6 7