Cloning the Ras Gene

Below you can see  the primers designed to prepare a PCR product containing the Ras gene flanked by attB recombination sites.

Forward attB
        T  S   L   Y   K   K   A   G
5'G GGG ACA AGT TTG TAC AAA AAA GCA GGC TTC GAA GGA GAT AGA ACC ATG ACA GAA TAC AAG CTT GTG GTG G 3'

Reverse attB
5'GGG GAC CAC TTT GTA CAA GAA AGC TGG GTC CTA GTG CTG CCG GAT CTC ACG CAC C 3'

c-H-Ras original sequence

  1 ATG ACA GAA TAC AAG CTT GTG GTG GTG GGC GCT GGA GGC GTG GGA AAG AGT
 52 GCC CTG ACC ATC CAG CTG ATC CAG AAC CAT TTT GTG GAC GAA TAC GAC CCC
102 ACT ATA GAG GAT TCC TAC CGG AAG CAG GTG GTC ATT GAT GGG GAG ACG TGC
153 CTG TTG GAC ATC CTG GAT ACC GCC GGC CTG GAG GAG TAC AGC GCC ATG CGG
204 GAC CAG TAC ATG CGC ACC GGG GAG GGC TTC CTG TGT GTG TTT GCC ATC AAC
255 AAC ACC AAG TCT TTT GAG GAC ATC CAC CAG TAC AGG GAG CAG ATC AAA CGG
306 GTG AAG GAC TCG GAT GAC GTG CCC ATG GTG CTG GTG GGG AAC AAG TGT GAC
357 CTG GCT GCA CGC ACT GTG GAA TCT CGG CAG GCT CAG GAC CTC GCC CGA AGC
408 TAC GGC ATC CCC TAC ATC GAG ACC TCG GCC AAG ACC CGG CAG GGA GTG GAG
459 GAT GCC TTC TAC ACG TTG GTG CGT GAG ATC CGG CAG CAC AAG CTG CGG AAG
510 CTG AAC CCT CCT GAT GAG AGT GGC CCC GGC TGC ATG AGC TGC AAG TGT GTG
561 CTC TCC TGA
 
Nucleotides 499-570 were not picked up by the PCR primers, instead we have inserted a stop codon. Thus the ras gene cloned here is a truncated mutant, and does not contain the amino acids 167-189.
You used these primers in a PCR reaction  and then ran the PCR products on a 1% agarose gel.


Click here to see your results.