A truncated form of the Carbon catabolite repressor 1 increases cellulase production in Trichoderma reesei

Mello-de-Sousa, Thiago M; Gorsche, Rita; Rassinger, Alice; Poças-Fonseca, Marcio J; Mach, Robert L; Mach-Aigner, Astrid R

doi:10.1186/s13068-014-0129-3

Biotechnology for Biofuels and Bioproducts

Table 2 Oligonucleotides used in this study

From: A truncated form of the Carbon catabolite repressor 1 increases cellulase production in Trichoderma reesei

Name	Sequence (5′ - 3′)	Usage
RG53	GAATTCAGATC	iv-FP, oligo-short
RG54	GCGGTGACCCGGGAGATCTGAATTC	iv-FP, oligo-long
RG89	[6-FAM]GTAGAGGCATGTTGTGAATCTGTGTCGGG	iv-FP, cbh1oligo3fw, EMSA
RG90	[6-FAM]GGTTGTATGCAAAACGCTCCGAGTCAGAC	iv-FP, cbh1oligo3rev, EMSA
RG215	CCAACGGCTTGTGGGGTTGCAGAAGCAACGGCAAAGCCCCACTTCCCCACGTTTGTTTCT^a	EMSA
RG216	AGAAACAAACGTGGGGAAGTGGGGCTTTGCCGTTGCTTCTGCAACCCCACAAGCCGTTGG
RG221	CCAACGGCTTGT T GGGTTGCAGAAGCAACGGCAAAGCCC A ACTTCCC A ACGTTTGTTTCT^b
RG222	AGAAACAAACGT T GGGAAGT T GGGCTTTGCCGTTGCTTCTGCAACCC A ACAAGCCGTTGG
RG178	TATCTCGAGTTTAGAAAAAAAAGCAGGT	pGEX-cre1-RG182 construction
RG182	ATTGGATCCATGCAACGAGCACAGTCTGCCGT	pGEX-cre1-RG182 construction
RG186	TTGAGTGCAGACGTGTGTGTAATCTT	Construction of QM6a-CREI₉₆
RG187	CCCTCCTTTGTTAGAAAAAAAAGCAGGTAATGG
RG188	TTTTTTCTAACAAAGGAGGGAGACGAGGTTG
RG189	CCTACATTGGATAACGGTGAGACTAGCGGCC
RG190	TCACCGTTATCCAATGTAGGTAAGTAGTAAGGG
RG191	GAATCAGTATTTTCTCATCTCCTTG
hph3′_fw	GACCTGCCTGAAACCGAACTG
hph5′_rev	GAAGAAGATGTTGGCGACCTCG
actfw	TGAGAGCGGTGGTATCCACG	qPCR
actrev	GGTACCACCAGACATGACAATGTTG
sar1fw	TGGATCGTCAACTGGTTCTACGA
sar1rev	GCATGTGTAGCAACGTGGTCTTT
cbh1f	GATGATGACTACGCCAACATGCTG
cbh1r	ACGGCACCGGGTGTGG
cbh2f	CTATGCCGGACAGTTTGTGGTG
cbh2r	GTCAGGCTCAATAACCAGGAGG
xyr1f	CCCATTCGGCGGAGGATCAG
xyr1r	CGAATTCTATACAATGGGCACATGGG
44747f	GCTCGAGCTGCAAGACAAGA
44747r	GCGGAGATCCATGAGCTTCT
epiactinTr_f	CTTCCCTCCTTTCCTCCCCCTCCAC	act CHART, region -226 to +24
epiactinTr_r	GCGACAGGTGCACGTACCCTCCATT	act CHART, region -226 to +24
episar1Tr_f	GTCAGGAAATGCCGCACAAGCAAGA	sar1 CHART, region -490 to -224
episar1Tr_r	TGTGTTTTACCGCCTTGGCCTTTGG	sar1 CHART, region -490 to -224
epicbh1_2Tr_f	GGATCGAACACACTGCTGCCTTTAC	cbh1 CHART, region -301 to -27
epicbh1_2Tr_r	GGTTTCTGTGCCTCAAAAGATGGTG	cbh1 CHART, region -301 to -27
epicbh2_2Tr_f	TGCAGCGCAACACTACACGCAACAT	cbh2 CHART, region -355 to -62
epicbh2_2Tr_r	TGCGCCTCATACAGGGTCACAGTCC	cbh2 CHART, region -355 to -62
epixyr1_2Tr_f	CCGACAGCAGCAGTAGTCAGGTTTT	xyr1 CHART, region -216 to +35
epixyr1_2Tr_r	TAGGCAGAATAGCGACGGAGAGGAT	xyr1 CHART, region -216 to +35

^aItalic letters indicate a CREI-binding site (5′-SYGGRG-3′).
^bBold letters indicate the introduced mutation in the CREI-binding site (5′-SYTGRG-3′).

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ISSN: 2731-3654

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