Expression and secretion of fungal endoglucanase II and chimeric cellobiohydrolase I in the oleaginous yeast Lipomyces starkeyi

Xu, Qi; Knoshaug, Eric P.; Wang, Wei; Alahuhta, Markus; Baker, John O.; Yang, Shihui; Vander Wall, Todd; Decker, Stephen R.; Himmel, Michael E.; Zhang, Min; Wei, Hui

doi:10.1186/s12934-017-0742-5

Table 1 Primers used in this study

From: Expression and secretion of fungal endoglucanase II and chimeric cellobiohydrolase I in the oleaginous yeast Lipomyces starkeyi

Gene cloning or construction	Primer	Primer nucleotide sequence
Plasmid backbone of pLS1	474_LS1_F1	AGTTTAGAGATGTACAAGGGGTATTGA
Plasmid backbone of pLS1	475_LS1_R1	GGCCAGAACGGCCGTGAGAATAGTAAAGGCGGTAGCGAGCTTCATGTTGGCTGTAGTGATACGGACGCAG
eg2 insertion to pLS1	476_LS1-F2	CTCGCTACCGCCTTTACTATTCTCACGGCCGTTCTGGCCCAACAGACGGTGTGGGGCCAA
	477_LS1-R2	CGCTGACAATGCACCTCAATACCCCTTGTACATCTCTAAACTTTATCACTTGCGGGCAAGGC
	505_EGII_F2	GAGCCCCATGACGTCAACATT
Plasmid backbone of pLS2	474_LS1_F1	AGTTTAGAGATGTACAAGGGGTATTGA
Plasmid backbone of pLS2	478_pLS2-R1	GCAATCTCGTCGAAGAATATAACGAGCAACCACAATAGGAGACAGTGATATCACTCCCAGAACAGCGATGAAAAAGTTGATCAGCAACATGTTGGCTGTAGTGATACGGACGCAG
eg2 insertion to pLS2	479_pLS2_F2	cctattgtggttgctcgttatattcttcgacgagattgcCAACAGACGGTGTGGGGCCAA
eg2 insertion to pLS2	477_LS1-R2	CGCTGACAATGCACCTCAATACCCCTTGTACATCTCTAAACTTTATCACTTGCGGGCAAGGC
Plasmid backbone of pLS3	474_LS1_F1	AGTTTAGAGATGTACAAGGGGTATTGA
Plasmid backbone of pLS3	480_LS3_R1	AGCTCCCAAAACCAACGTCGCCAACAGAATGTTAACCAGTACAATCCGTGTGATTGAGGGGACCATTGTAAATATTGAAGGCACGTAGATTAATGTCATGTTGGCTGTAGTGATACGGACGCAG
eg2 insertion to pLS3	481_LS3_F1	ctggttaacattctgttggcgacgttggttttgggagctCAACAGACGGTGTGGGGCCAA
eg2 insertion to pLS3	477_LS1-R2	CGCTGACAATGCACCTCAATACCCCTTGTACATCTCTAAACTTTATCACTTGCGGGCAAGGC
Plasmid backbone of pLS4	474_LS1_F1	AGTTTAGAGATGTACAAGGGGTATTGA
Plasmid backbone of pLS4	475_LS1_R1	GGCCAGAACGGCCGTGAGAATAGTAAAGGCGGTAGCGAGCTTCATGTTGGCTGTAGTGATACGGACGCAG
cbh1 insertion to pLS4	482_pLS4_F1	GCTCGCTACCGCCTTTACTATTCTCACGGCCGTTCTGGCCCAGCAGGCAGGGACCGCTACC
cbh1 insertion to pLS4	483_pLS4_R1	GCTGACAATGCACCTCAATACCCCTTGTACATCTCTAAACTTTATCACAGGCATTGGGAGTAGTAAGGG
Plasmid backbone of pLS5	474_LS1_F1	AGTTTAGAGATGTACAAGGGGTATTGA
Plasmid backbone of pLS5	478_pLS2-R1	GCAATCTCGTCGAAGAATATAACGAGCAACCACAATAGGAGACAGTGATATCACTCCCAGAACAGCGATGAAAAAGTTGATCAGCAACATGTTGGCTGTAGTGATACGGACGCAG
cbh1 insertion to pLS5	484_LS5_F1	cctattgtggttgctcgttatattcttcgacgagattgcCAGCAGGCAGGGACCGCTACC
cbh1 insertion to pLS5	483_pLS4_R1	GCTGACAATGCACCTCAATACCCCTTGTACATCTCTAAACTTTATCACAGGCATTGGGAGTAGTAAGGG
Plasmid backbone of pLS6	474_LS1_F1	AGTTTAGAGATGTACAAGGGGTATTGA
Plasmid backbone of pLS6	480_LS3_R1	AGCTCCCAAAACCAACGTCGCCAACAGAATGTTAACCAGTACAATCCGTGTGATTGAGGGGACCATTGTAAATATTGAAGGCACGTAGATTAATGTCATGTTGGCTGTAGTGATACGGACGCAG
cbh1 insertion to pLS6	485_LS6_F1	ctggttaacattctgttggcgacgttggttttgggagctCAGCAGGCAGGGACCGCTACC
cbh1 insertion to pLS6	483_pLS4_R1	GCTGACAATGCACCTCAATACCCCTTGTACATCTCTAAACTTTATCACAGGCATTGGGAGTAGTAAGGG
Plasmid backbone of pLS7	474_LS1_F1	AGTTTAGAGATGTACAAGGGGTATTGA
Plasmid backbone of pLS7	486_LS7_R1	TGCAGCTCCCAAAACCAACGTCGCCAACAGAATGTTAACCAGTACAATCCGTGTGATTGAGGGGACCATGTTGGCTGTAGTGATACGGACGCAG
eg2 insertion to pLS7	487_LS7_F1	ggttaacattctgttggcgacgttggttttgggagctgcaCAACAGACGGTGTGGGGCCAA
eg2 insertion to pLS7	477_LS1-R2	CGCTGACAATGCACCTCAATACCCCTTGTACATCTCTAAACTTTATCACTTGCGGGCAAGGC
Plasmid backbone of pLS8	474_LS1_F1	AGTTTAGAGATGTACAAGGGGTATTGA
Plasmid backbone of pLS8	486_LS7_R1	TGCAGCTCCCAAAACCAACGTCGCCAACAGAATGTTAACCAGTACAATCCGTGTGATTGAGGGGACCATGTTGGCTGTAGTGATACGGACGCAG
cbh1 insertion to pLS8	488_LS8_F1	ggttaacattctgttggcgacgttggttttgggagctgcaCAGCAGGCAGGGACCGCTACC
cbh1 insertion to pLS8	483_pLS4_R1	GCTGACAATGCACCTCAATACCCCTTGTACATCTCTAAACTTTATCACAGGCATTGGGAGTAGTAAGGG
Correct insertion sequencing	491_ES_F1	GTTGAGCTGCCGCGTGTTCTG
	492_ES_F2	CACACGCGGCTAACACCGATT
	493_ES_R1	CGTTCGATTGGGTTGGTTGGAC
	494_ES_R2	CTGCCACGTCGTTCGATTGGG
eg2 sequencing	497_EGII_F1	CAACAGACGGTGTGGGGCCAA
	498_EGII_R1	TTATCACTTGCGGGCAAGGC
	505_EGII_F2	GAGCCCCATGACGTCAACATT
cbh1 sequencing	499_CBHI_F1	CAGCAGGCAGGGACCGCTACC
	500_CBHI_R1	TTATCACAGGCATTGGGAGTAGTAAGGG
	506_CBHI_F2	ATACTGGAATCGGCGACCAC

pLS1, 2, 3, 4, 5, 6, 7, and 8 are the plasmids constructed and used in this study (also see Table 3)

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ISSN: 1475-2859

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