Expression of a gene for a modified green-fluorescent protein

940204 RV9 36 RUTG 3.0-017

FIGURE 1

The amino acid sequence of a pre-coelenterazine peptide:

Xaa Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro lie Leu Val 1 5 10 15

Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly Glu 20 25 30

Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe lie Cys 35 40 45

Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Phe

50 55 60

Tyr Tyr Gly Val Gin Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gin 65 70 75 80

His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gin Glu Arg 85 90 95

Thr lie Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val 100 105 110

Lys Phe Glu Gly Asp Thr Leu Val Asn Arg lie Glu Leu Lys Gly lie 115 120 125

Asp Phe Lys Glu Asp Gly Asn lie Leu Gly His Lys Leu Glu Tyr Asn 130 135 140

Tyr Asn Ser His Asn Val Tyr lie Met Ala Asp Lys Gin Lys Asn Gly 145 150 155 160

lie Lys Val Asn Phe Lys lie Arg His Asn lie Glu Asp Gly Ser Val 165 170 175

Gin Leu Ala Asp His Tyr Gin Gin Asn Thr Pro He Gly Asp Gly Pro 180 185 190

Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gin Ser Ala Leu Ser 195 200 205

Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val

210 215 220

Thr Ala Ala Gly He Thr His Gly Met Asp Glu Leu Tyr Lys 225 230 235

U.S. Patent

Apr. 21, 1998 Sheet 2 of 3

5,741,668

FIG. 2

The nucleotide sequence of cDNA segment cjfp.(C197A) encoding the pre-coelenterazine peptide of Figure 1:

ATGGCTAGCA AAGGAGAAGA ACTTTTCACT GGAGTTGTCC CAATTCTTGT TGAATTAGAT 60

GGTGATGTTA ATGGGCACAA ATTTTCTGTC AGTGGAGAGG GTGAAGGTGA TGCAACATAC 120

GGAAAACTTA CCCTTAAATT TATTTGCACT ACTGGAAAAC TACCTGTTCC ATGGCCAACA 180

CTTGTCACTA CTTTCTATTA TGGTGTTCAA TGCTTTTCAA GATACCCAGA TCATATGAAA 240

CAGCATGACT TTTTCAAGAG TGCCATGCCC GAAGGTTATG TACAGGAAAG AACTATATTT 300

TTCAAAGATG ACGGGAACTA CAAGACACGT GCTGAAGTCA AGTTTGAAGG TGATACCCTT 360

GTTAATAGAA TCGAGTTAAA AGGTATTGAT TTTAAAGAAG ATGGAAACAT TCTTGGACAC 4 20

AAATTGGAAT ACAACTATAA CTCACACAAT GTATACATCA TGGCAGACAA ACAAAAGAAT 480

GGAATCAAAG TTAACTTCAA AATTAGACAC AACATTGAAG ATGGAAGCGT TCAACTAGCA 540

GACCATTATC AACAAAATAC TCCAATTGGC GATGGCCCTG TCCTTTTACC AGACAACCAT 600

TACCTGTCCA CACAATCTGC CCTTTCGAAA GATCCCAACG AAAAGAGAGA CCACATGGTC 660

CTTCTTGAGT TTGTAACAGC TGCTGGGATT ACACATGGCA TGGATGAACT ATACAAATAA 720

ATGTCCAGAC TTCCAATTGA CACTAAAGTG TCCGAACAAT TACTAAAATC TCAGGGTTCC 780

TGGTTAAATT CAGGCTGAGA TATTATTTAT ATATTTATAG ATTCATTAAA ATTGTATGAA 840

TAATTTATTG ATGTTATTGA TAGAGGTTAT TTTCTTATTA AACAGGCTAC TTGGAGTGTA 900

TTCTTAATTC TATATTAATT ACAATTTGAT TTGACTTGCT CAAA 944