Figure 2 of
Fautsch, Mol Vis 2004;
10:417-425.Figure 2 of
Fautsch, Mol Vis 2004;
10:417-425.
Figure 2. Oligonucleotides used to create myocilin constructs
Oligonucleotide sequence, specific nucleotide location, oligonucleotides used to amplify 5' and 3' intermediate PCR fragments, and fragment lengths are listed for templates used. Highlighted base pairs represent alanine codon substitution. Asterisks indicate complementary strand. A: In vitro studies. B: In vivo studies.
A:
Construct Oligo
name name Sequence Nucleotide
--------- ------- ------------------------------------------- ----------
C1 Cy1-5al 5'-AGAGTGGCCGAGCCCAGTATACCTTCAGTG-3' 164- 193
Cy1-3al 5'-AGGTATACTGGGCTCGGCCACTCTGGTCATTG-3' 156- 187*
C2 Cy2-5al 5'-GAATCCAGCGCCCCAGAGCAGAGCCAGGCCATG-3' 208- 240
Cy2-3al 5'-GCTCTGCTCTGGGGCGCTGGATTCATTGGGACTG-3' 198- 226*
C3 Cy3-5al 5'-AGGGGCCAGGCTCCCCAGACCCGAGACACTG-3' 580- 610
Cy3-3al 5'-TCTGGGGAGCCTGGCCCCTTCTCAGCCTTGC-3' 568- 598*
C4 Cy4-5al 5'-ACACCGGAGCTGGAGAACTAGTTTGGGTAG-3' 761- 790
Cy4-3al 5'-CTAGTTCTCCAGCTCCGGTGTCTCCCTCTCCAC-3' 749- 781*
C5 Cy5-5al 5'-TTCATCATCGCTGGCACCTTGTACACCGTCAGC-3' 1324-1359
Cy5-3al 5'-CAAGGTGCCAGCGATGATGAAGGCATTGGCGAC-3' 1312-1344*
MYOC-5' 5'-ATGCGAATTCAGGACAGCTCAGCTCAGGAAGGC-3' 133- 155
MYOC-3' 5'-ATGCGGATCCTTATCACATCTTGGAGAGCTTGATGTC-3' 1528-1551*
Construct Intermediate Amplified Fragment
name PCR product Oligonucleotide pair nucleotides length
--------- ------------ -------------------- ----------- --------
C1 5' fragment MYOC-5', Cy1-3al 133- 187 55
3' fragment MYOC-3', Cy1-5al 164-1551 1388
C2 5' fragment MYOC-5', Cy2-3al 133- 226 94
3' fragment MYOC-3', Cy2-5al 208-1551 1344
C3 5' fragment MYOC-5', Cy3-3al 133- 598 466
3' fragment MYOC-3', Cy3-5al 580-1551 972
C4 5' fragment MYOC-5', Cy4-3al 133- 781 649
3' fragment MYOC-3', Cy4-5al 761-1551 791
C5 5' fragment MYOC-5', Cy5-3al 133-1344 1212
3' fragment MYOC-3', Cy5-5al 1359-1551 193
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B:
Construct Oligo
name name Sequence Nucleotide
--------- --------- -------------------------------------------- ----------
Arg82Cys Arg5-3cy 5'-AGCAGCACCCAATGCTTAGACCTGGAGGCCAC-3' 268- 299
Arg3-5cy 5'-CTCCAGGTCTAAGCATTGGGTGCTGCTGTCTC-3' 263- 293*
Cys433Arg Cys433-5' 5'-GCCTTCATCATCCGTGGCACCTTGTACACCGTCAGC-3' 1321-1356
Cys433-3' 5'-GGTGCCACGGATGATGAAGGCATTGGCGAC-3' 1312-1341*
MYOC5-H3 5'-TAGCGAAGCTTTCTGCAATGAGGTTCTTCTGTGC-3' 31- 53
MYOC-Stop 5'-ATGCGTCTAGATCATCACATCTTGGAGAGCTTGATGTC-3' 1528-1551*
Construct Intermediate Amplified Fragment
name PCR product Oligonucleotide pair nucleotides length
--------- ------------ -------------------- ----------- --------
Arg82Cys 5' fragment MYOC5-H3, Arg3-5cy 31- 293 263
3' fragment MYOC-Stop, Arg5-3cy 268-1551 1284
Cys433Arg 5' fragment MYOC5-H3, Cys433-3' 31-1341 1311
3' fragment MYOC-Stop, Cys433-5' 1321-1551 231
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