Autolysis of Pichia pastoris induced by cold

AMB Express

Table 2 Primers used in this study

Primer^a	Sequence (5′→3′)^b	Restriction enzyme	Underlined sequence^c
YL1-F	GGGGtcgcgaGAATCTTCCTCAGAAGAGGAAT	NruI	Homologous Pcctα region corresponding to nucleotides (nt) 886842 to 886863
YL1-R	GCGCggatccTCACCTCCTAGGAAAAGGGTAT	BamHI	Homologous Pcctα region corresponding to nt 887263 to 887284
YL2-F	CCCCagatctCCCAAATGTCATTCTCTTCCAA	BglII	Homologous eng region corresponding to nt 802095 to 802116
YL2-R	GCGCctcgagTTAAACGGAATTAGCCAGTCCT	XhoI	Homologous eng region corresponding to nt 805153 to 805132
YL3-F	GCGCgtcgacTGAGTTTTAGCCTTAGACATGACT	SalI	Homologous aox1 TT region corresponding to nt 627 to 650
YL3-R	GGGGcacgtgCCGCACAAACGAAGGTCTCA	PmlI	Homologous aox1 TT region corresponding to nt 970 to 957
YL4-F	GGGGatttaaatCCCACACACCATAGCTTCAAA	SwaI	Homologous Ptef1 region corresponding to nt 971 to 993
YL4-R	GGGGcacgtgTCTCCAGCTTGCAAATTAAAG	PmlI	Homologous cyc1 TT region corresponding to nt 2142 to 2122
YL5-F	GGGGtcgcgaGTATATTGAGGAGCAGAACTAG	NruI	Homologous leu2 region corresponding to nt 86482 to 86503
YL5-R	GGGGcacgtgCTTTTTCCGAACCTTACAGTAG	PmlI	Homologous leu2 region corresponding to nt 87943 to 87922

^a F forward primer, R reverse primer
^bLowercase letters indicates the restriction endonuclease recognized site denoted in the next column
^cNucleotide numbering has been taken according to the sequence depicted in the access number mentioned in the section of materials and methods