Development of the Multiple Gene Knockout System with One-Step PCR in Thermoacidophilic Crenarchaeon Sulfolobus acidocaldarius

Table 2

Primers used in this study.

Primer	Sequence^a (5-3)
SSOlacS-F	TTTCTGCAGTGTTTTTCTCTATATCAATCTC
SSOlacS-R	TTTGGATCCATCTAAATGACTTTCCAATTAG
E800-F	ACTTCTCCTCCTTATTATAACG
E800-R	GGATTCTCTTACTTTTCTAAAG
E50-F	TGAGGGAAAAAATAAACGAAAAG
E50-R	GACCTTGAATTTGAAGTGGC
E40-F	AATAAACGAAAAGTTAGAAAAGAA
E40-R	TTGAAGTGGCGTCTCTAGAT
E30-F	AAGTTAGAAAAGAAATCTCAG
E30-R	GTCTCTAGATCGTTAGCAC
E20-F	AGAAATCTCAGTGACTGCAG
E20-R	CGTTAGCACATAAAGTCAGTC
E10-2F	ATCTCAGTGATGTTTTTCTCTATATCAATCTC
E10-2R	TAAAGTCAGTACTCCTAGATCTAAAACTAAAG
phr-pop-F	ATGATGTTTTAGAACAAAAGATTATTTAGTTGTAGTATAATCATTAGTCCAAGGGGAAAAAGTATAAAGAGAAAACAATGTTTTTCTCTATATCAATCTC
phr-pop-R	ATTATCGAATAATCTCAAATCCCTTCTAAATATTACTGCACAATCCACACTCCTAGATCTAAAACTAAAG
phr-out-F	AACGCTGGGATGCTGATAAG
phr-out-R	ATGACCAGACTACTAACGTAC
argD-pop-F	AAACCTAAACGTCATCAAATGTTTTTCGCAGATAGAAGTTCAGAGTGATGTTTTTCTCTATATCAATCTC
argD-pop-R	AATAGGTAGATGATGAAATTAAAAAAAGAGATCGATGATCAACTCAGCCTCCTTTTCTATTACCCTCCATCACCACTTACTCCTAGATCTAAAACTAAAG
argD-F-F	TTACTTTATATATCTCATTCTG
argD-F-R	CTAATTAGGGAAATTGGTTAC
SsoargD-KpnI-F	TTTGGTACCCTTATTACCTAGATATAACGTT
SsoargD-PstI-R	TTTGTCGACTACTGCTTTGATCAAATATAAG

^aRestriction sites are underlined and sequences of MONSTER primers that anneal with the pyrE-lacS marker genes are in bold.