Skip to main content

Advertisement

Table 1 Hprt sequences

From: Simultaneous targeted exchange of two nucleotides by single-stranded oligonucleotides clusters within a region of about fourteen nucleotides

Sequences of coding strand of hprt intron 2 and exon 3 in V79 wildtype and V79-151 cells.
V79 wildtype position 151
DNA sequence 5'-ttgtag G ACT GAA AGA CTT GCC C GA GAT GTC ATG AAA GAG ATG GGA-3
amino acids Thr Glu Arg Leu Ala Arg Asp Val Met Lys Glu Met Gly
V79-151 position 151
DNA sequence 5'-ttgtag G ACT GAA AGA CTT GCC T GA GAT GTC ATG AAA GAG ATG GGA-3
amino acids Thr Glu Arg Leu Ala Stop Asp Val Met Lys Glu Met Gly
Positions of the intended first nucleotide exchanges for correcting the hprt mutation.
  Exchange of nucleotide on position 151
DNA sequence 5'-ttgtag G ACT GAA AGC CTT GCC C GA GAT GTT ATG AAA GAG ATG GGA-3'
amino acids Thr Glu Arg Leu Ala Arg Asp Val Met Lys Glu Met Gly
  Exchange of nucleotide on position 153
DNA sequence 5'-ttgtag G ACT GAA AGC CTT GCC TGC GAT GTT ATG AAA GAG ATG GGA-3'
amino acids Thr Glu Arg Leu Ala Cys Asp Val Met Lys Glu Met Gly
Positions of the intended second nucleotide exchange generating a silent mutation
Position Base exchange Sequence of coding strand of hprt intron 2 in V79-151 cells.
141 A to G 5'-ttgtag G ACT GAG AGA CTT GCC T GA GAT GTC ATG AAA GAG ATG GGA-3
144 A to G 5'-ttgtag G ACT GAA AGG CTT GCC T GA GAT GTC ATG AAA GAG ATG GGA-3
147 T to C 5'-ttgtag G ACT GAA AGA CTC GCC T GA GAT GTC ATG AAA GAG ATG GGA-3
150 C to T 5'-ttgtag G ACT GAA AGA CTT GCT T GA GAT GTC ATG AAA GAG ATG GGA-3
156 T to C 5'-ttgtag G ACT GAA AGA CTT GCC T GA GAC GTC ATG AAA GAG ATG GGA-3
159 C to T 5'-ttgtag G ACT GAA AGA CTT GCC T GA GAT GTT ATG AAA GAG ATG GGA-3
165 A to G 5'-ttgtag G ACT GAA AGA CTT GCC T GA GAT GTC ATG AAG GAG ATG GGA-3
168 G to A 5'-ttgtag G ACT GAA AGA CTT GCC T GA GAT GTC ATG AAA GAA ATG GGA-3
  1. The table shows the sequence of coding strand of hprt intron 2 (lower case letters) and exon 3 (upper case letters) in V79 wildtype cells and V79-151 cells, which carry a stop codon at position 151. Also shown is the sequence of coding strand of hprt after base exchange at position 151 or 153 and after base exchange at the second mismatch position, which generates a silent mutation. The exchanged bases are indicated in bold. The respective second base exchanges are generated through the following oligonucleotides: position 141: oligonucleotide H7-151-141; position 144: H5-151-144; position 147: H3-151-147, O45-151-147, C153-147; position 150: H2-151-150, O45-151-150, C153-150; position 156: H4-151-156, O42-151-156, C153-156; position 159: H6-151-159, O25-151-159, O30-151-159, O45-151-159, O48-151-159, C153-159; position 165: H8-151-165; position 168: H9-151-168.