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Table 2 Primers used for PCR

From: Cloning of a novel thermostable glucoamylase from thermophilic fungus Rhizomucor pusillus and high-level co-expression with α-amylase in Pichia pastoris

Primers

Function

Sequence (5’-3’)

R.pGJf

Primers for conserved regions of glucoamylase gene

TGGGGHMGHCCNCARAATGAYGG

R.pGJr

RTCGTCAGGRTANCKRCCRATDGC

5-1SP1

Primers for 5’-flanking region of glucoamylase gene

TCTTCCCTAGAATTGATGCGTGTGA

5-1SP2

AGTCTAAATCCTTGAATATCGCCGG

5-1SP3

AAGGATAAAGGTCGATGCACGCAGT

5-2SP1

Primers for 5’-flanking region of glucoamylase gene

ATCCTCCTCCTCCCATGAAGAAACA

5-2SP2

AGCAATATGTTGGTTGCGGTTGATC

5-2SP3

GAAAAGCATCAGCAGCACCTGAATC

3-1SP1

Primers for 3’-flanking region of glucoamylase gene

TGGATGTTTCTATCCTATTGGCAGC

3-2SP2

CCACATAAATATGGATCCATTGCCG

3-3SP3

TTGTATCCACTCAATCGGGAGCAGC

R.pGWr

Primers for glucoamylase gene glu’

GATGAAAGCAGCGTACGACCATGTC

R.pGWf

TGTGCAAGAATCTACCCTTTTCGAG

R.pGf1

Primers containing putative start codon for cDNA of glucoamylase gene

ATGCGTTATGCAACCCCGC

R.pGf2

ATGCTCTTCGCTTTTTGCTATTGT

R.pGf3

ATGTCTTACCGGAAGCAATTTCT

R.pGf4

ATGGGAGGAGGAGGATCTTGGT

R.pGr1

Primers containing putative stop codon for cDNA of glucoamylase gene

GGCGTTATTTATTACCCTCTTTTGACC

R.pGf

Primers for cDNA of glucoamylase gene

ATGGACACGCGATTCAGCCC

R.pGr

AGCGTACGACCATGTCAGGTCG

R.pGEcoRf

Primers containing restriction enzyme sites for cDNA of glucoamylase gene

GGAATTCATGCGTTATGCAACCCCGC

R.pGNotr

TTGCGGCCGCTTACCCTCTTTTGACCA

R.pAf

Primers for cDNA of α-amylase gene

ATGAAATTCAGCATCTCTCTCTCGG

R.pAr

TTAAGCAGAGGTGAAGATAGCGGA

R.pAEf

Primers containing restriction enzyme sites for cDNA of α-amylase gene

GAATTCAGCCCTTTGCCCCAACAGCA

R.pANr

TTGCGGCCGCTTAAGCAGAGGTGAAGATAG

  1. The primers are denoted as follows: the start codon and stop codon are underlined; the cons`ervative codons are boxed; the dotted line indicated restriction enzyme sites.
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BMC Biotechnology

ISSN: 1472-6750

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