Table 2 Primers used for the construction of different FAK and Src expression vectors.

From: Simultaneous deactivation of FAK and Src improves the pathology of hypertrophic scar

Target sequence/mutation

Accession no.

Primer orientation

Primer sequence (5′ → 3′)

Nucleotide position

Primers for the amplification of full-length human FAK and Src

FAK

L13616.1

S

ACGGTACCATGGCAGCTGCTTACCTTGA

233–252

 

L13616.1

AS

ACCTCGAGTCAGTGTGGTCTCGTCTGCC

3372–3391

Src

NM_005417.4

S

ACGGTACCATGGGTAGCAACAAGAGCAA

450–469

 

NM_005417.4

AS

ACCTCGAGCTAGAGGTTCACCCCGGGCT

2041–2060

Mutagenic primers used during three-step mutagenic PCR of FAK or Src

FAK Tyr407 → Phe

L13616.1

S

TATAGATGAAGAAGATACTTTTACCATGCCCTCAACCAGGG

1432–1472

 

L13616.1

AS

CCCTGGTTGAGGGCATGGTAAAAGTATCTTCTTCATCTATA

1432–1472

Src Tyr529 → Glu

NM_005417.4

S

CACGTCCACCGAGCCCCAGGAACAGCCCGGGGAGAACCTCT

2018–2058

 

NM_005417.4

AS

AGAGGTTCTCCCCGGGCTGTTCCTGGGGCTCGGTGGACGTG

2018–2058

Flanking primers used during three-step mutagenic PCR of FAK or Src

pcDNA3.1(+)

EF550208.1

S

TAATACGACTCACTATAGGGAGA

863–885

FAK

L13616.1

AS

ACCTCGAGTCAGTGTGGTCTCGTCTGCC

3372–3391

Src

NM_005417.4

AS

ACCTCGAGCTAGAGGTTCACCCCGGGCT

2041–2060

  1. Sequences marked by dotted underlining represent the restriction enzyme recognition sites (GGTACC for Kpn I; CTCGAG for Xho I) added for cloning into pcDNA3.1(+) Mammalian Expression Vector. Sequences marked by solid underlining represent sites of mutation on FAK or Src. S, sense; AS, antisense. ATG, start codon; TCA or CTA, stop codons.
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