Data for the Constructed Mutant: YFHEdd

General Information

Name of the Mutant	YFHEdd (deletion (double-cross))
Host	168
Used Vector	pMutin2
Used Enzyme Sites	XhoI-BamHI & EcoRI-XhoI
Gene Size	108 (nt)

Growth & Expression (reporter lacZ):

in DSM:	yes
in MM:	no

Mutant Submission to NIG:	yes
Correspondence:

[Construction of the Mutant]

Primers used for Deletion: Upstream: Forward Primer: (ctcctcga tag) yfhE-UF: ctcctcgaGCGGAACGATCGGAACG Reverse Primer: (ggagga tag) yfhE-UBR: ggaggaTCCATAAAAAGGACCTCC Downstream: Forward Primer: (gcggaattc tag) yfhE-DEF: gcggaattcAAAGCTGCTAAGCGGAAG Reverse Primer: (ctcctcga tag) yfhE-DR: ctcctcgaGGATAATGAGCGTTTCGC
Linearization:
Length of the Deletion: 6 to 90 Length: 85

Forward Primer:	(ctcctcga tag)	yfhE-UF:	`ctcctcgaGCGGAACGATCGGAACG`
Reverse Primer:	(ggagga tag)	yfhE-UBR:	`ggaggaTCCATAAAAAGGACCTCC`

Forward Primer:	(gcggaattc tag)	yfhE-DEF:	`gcggaattcAAAGCTGCTAAGCGGAAG`
Reverse Primer:	(ctcctcga tag)	yfhE-DR:	`ctcctcgaGGATAATGAGCGTTTCGC`

[Confirmation of the Constructed Mutant]

Primer sets used for the check

No. Primer set Length of PCR product

Forward Primer	hGBR	`gcgcggatccGTAATCGTTAAGCCTATGG`
Reverse Primer	hBHF	`gccgaagcttAAGCATTTACAAGCAGTCC`

Forward Primer	yfhE-UF	`ctcctcgaGCGGAACGATCGGAACG`
Reverse Primer	yfhE-DR	`ctcctcgaGGATAATGAGCGTTTCGC`

III

Forward Primer	yfhE-UBR	`ggaggaTCCATAAAAAGGACCTCC`
Reverse Primer	yfhE-DEF	`gcggaattcAAAGCTGCTAAGCGGAAG`

Digestion of the PCR product: no

Enzyme used:

Figure

Comment: ;.;.

1st Level Phenotype Analysis of YFHEdd