Skip to main content

Table 1 List of microorganisms and plasmids used in this study

From: Cloning and expression of an active aspartic proteinase from Mucor circinelloides in Pichia pastoris

Strain or plasmid

Genotype

Reference

Strains

  

Escherichia coli

TOP10

F- mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu) 7697 galU galK rpsL (StrR) endA1 nupG

10

Pichia pastoris

  

X-33

Wild type

Invitrogen

Mucor circinelloides

  

DSM 2183

Wild type

German resource centre for biological material

Plasmids

  

pGAPZα-A

The pGAPZα-A vector use the GAP promoter to constitutively express recombinant proteins in Pichia pastoris. Contains the zeocin resistance gene (Sh ble).

Invitrogen

pGAPZα+MCAP

pGAPZα-A derivative carrying the whole MCAP gene1.

This work

pGAPZα+MCAP-2

pGAPZα-A derivative carrying the MCAP gene without an intron1.

This work

pGAPZα+MCAP-3

pGAPZα-A derivative carrying the MCAP gene without an intron2.

This work

pGAPZα+MCAP-5

pGAPZα-A derivative carrying the MCAP gene without a signal sequence and without an intron2.

This work

pGAPZα+MCAP SP-1

pGAPZα-A derivative carrying from the amino acid sequence number 67 to 394 of the MCAP gene without an intron1.

This work

pGAPZα+SyMCAP-6

pGAPZα-A derivative carrying the MCAP gene without signal sequence and without intron. The original MCAP gene was adapted to the optimal codon usage of P. pastoris. The insert was cloned flush with the Kex2 cleavage site and in frame of the α- factor signal sequence and in frame with the C-terminal polyhistidine tag into the XhoI and NotI site of the pGAPZα-A.

This work

  1. 1The insert was cloned in frame with the α-factor signal sequence and in frame with the C-terminal polyhistidine tag into the EcoRI and NotI sites of the pGAPZα-A.
  2. 2The insert was cloned flush with the Kex2 cleavage site and in a frame of the α-factor signal sequence and in a frame with the C-terminal polyhistidine tag into the XhoI and NotI site of the pGAPZα-A.