Increased isobutanol production in Saccharomyces cerevisiae by overexpression of genes in valine metabolism

Table 4 Primer sequences

Fragment	Primer	Primer sequence (5' to 3')	Restriction site
PGK1 promoter	Forward	AAAAAACCCGGG TCTAACTGATCTATCCAAAACTG	Xma l
	Reverse	TGTTTTATATTTGTTGTAAAA AGTAGA
ILV2	Forward	TCTACTTTTTACAACAAATATAAAACA ATGATCAGACAATCTACGCTAA
	Reverse	AAAAAAGGCGCC CAAGCTTGCAATTTTTGACG	Nar I
ILV3	Forward	TCTACTTTTTACAACAAATATAAAACA ATGGGCTTGTTAACGAAAGT
	Reverse	AAAAAAGGCGCC CTTTGGTAGAGGTGGCTTCG	Nar I
ILV5	Forward	TCTACTTTTTACAACAAATATAAAACA ATGTTGAGAACTCAAGCCGC
	Reverse	AAAAAAGGCGCC ATTCGCGTTTCGGTTCTTGT	Nar I
ILV6	Forward	TCTACTTTTTACAACAAATATAAAACA ATGCTGAGATCGTTATTGCA
	Reverse	AAAAAAGACGTC AACATCCCAATATCCGTCCA	Aat II
BAT2	Forward	TCTACTTTTTACAACAAATATAAAACA ATGACCTTGGCACCCCTAGA
	Reverse	AAAAAAGGCGCC GAATTGTCTTGAGTTGCTTCTAAGGTA	Nar I
qILV2	Forward	TCCAAGGTTGCCAACGACACAG
	Reverse	TGTTGAGCAGCCCACATTTGATG
qILV3	Forward	TTGCACCTCCACCTCGTTACAC
	Reverse	ACCGTTGGAAGCGTTGGAAACC
qILV5	Forward	TTACGCCGTCTGGAACGATGTC
	Reverse	GAACCAATGGCAACGGCCAAAG
qILV6	Forward	TACCATGGTGCGTTGCAGTTCC
	Reverse	AGGTCTTGTTGCGTGTCTGTGC
qBAT2	Forward	GAAATCGGCTGGAAAGGCGAAC
	Reverse	CTTTGGCCAATGGACCGGTTTG
qACT1	Forward	TGGATTCTGAGGTTGCTGCTTTGG
	Reverse	ACCTTGGTGTCTTGGTCTACCG

^aRestriction sites used for cloning are shown in bold. The reverse and complementary sequences used for fusion PCRs are shown in italics. The primers for qILV2 qILV3 qILV5 qILV6 qBAT2 and qACT1 amplifications were used for quantitative real-time polymerase chain reactions.

ISSN: 2731-3654