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Table 4 Plasmids, strains and primers used

From: Efficient one-step production of (S)-1-phenyl-1,2-ethanediol from (R)-enantiomer plus NAD+–NADPH in-situ regeneration using engineered Escherichia coli

Plasmids, strains, primers

Description

Sources

Plasmids

  

pMD18-T

2.7 kb, Ampr

Takara

pMD18-RCR

3.7 kb, pMD18-T containing rcr, Ampr

Novagen

pMD18-SCR

3.5 kb, pMD18-T containing scr, Ampr

This work

pMD-PntA

4.2 kb, pMD18-T containing PntA, Ampr

This work

pMD-PntB

4.1 kb, pMD18-T containing PntB, Ampr

This work

pETDuet™-1

5.4 kb, contains two multiple cloning sites, Ampr

Novagen

pACYCDuet™-1

4.0 kb, contains two multiple cloning sites, Cm r

Novagen

pET-RS

7.4 kb, pETDuet™-1 containing rcr and scr, Ampr

This work

pACYC-AB

6.8 kb, pACYCDuet™-1 containing PntA and PntB, Cm r

This work

Strains

  

C. parapsilosis CCTCC M203011

DNA donors of rcr and scr genes

This laboratory

E. coli K12

DNA donors of PntA and PntB genes

This laboratory

E. coli JM109

recA 1 supE 44 endA 1 hsdR 17 gyrA 96 relA 1 thi(Lac-proAB)F’

This laboratory

E. coli BL21(DE3)

F-ompT hsdS B (r B -m B -) gal dcm (DE3)

Novagen

CK

E. coli BL21 bearing pETDuet™-1 and pACYCDuet™-1

 

RS

E. coli BL21 bearing pET-RS

This work

AB

E. coli BL21 bearing pACYC-AB

This work

RSAB

E. coli BL21 bearing pET-RS and pACYC-AB

This work

Primers

5′ → 3′

 

RCR_F

ATCGATCGCATATG TCAATTCCATCAAGCCAGTACGG(NdeI)

This work

RCR_R

TGACTCTCGAG CTATGGATTAAAAACAACACGACC(Xho I)

This work

SCR_F

ATCGAATTC GATGGGCGAAATCGAATCTTATTG(Eco RI)

This work

SCR_R

TGACTGCGGCCGC CTATGGACACGTGTATCCACCGTC(Not I)

This work

PntA_F

CGCGGATCCATGCGAATTGGCATACCAAG (Bam HI)

This work

PntA_R

CCCAAGCTT TTAATTTTTGCGGAACATTTTC (Hin dIII)

This work

PntB_F

CGCGATATC ATGTCTGGAGGATTAGTTAC (Eco RV)

This work

PntB_R

CCCCTCGAG TTACAGAGCTTTCAGGATTG (Xho I)

This work