Table 1 Plasmids and oligonucleotides used in this work

pFC

Psychrophilic gene-expression vector, containing the T/R box, the promoter and termination region of the P. haloplanktis TAC125 asp C gene and the chloramphenicol resistance gene

[6]

pFCamy ΔCt

pFC containing a truncated version of the amy gene devoid of the C-terminal propeptide encoding portion

[6]

pFCamy ΔCt-blaM

pFC containing the amy ΔCt-blaM gene encoding the chimerical protein Amy-BlaM

[6]

pVS

pGEM7Z vector containing a conjugation transfer origin (oriT), the counter selectable marker pheS ^Gly294 and the ampicillin resistance gene

This work

Oligonucleotides

Oligo oriTEco RIfw

5'-TTGAATTCTCGCACGATATACAGG-3'

Oligo oriTSac Irv

5'-AAGAGCTCTTGAAGACGAAAGGG-3'

Oligo gspESph Ifw

5'-TTGCATGCATGCGCATCATCCGG-3'

Oligo gspESac Irv

5'-AAGAGCTCCAATATCGAGCTTAGCC-3'

Oligo gspESac Ifw

5'-TTGAGCTCCTAAAGTAGGTATGACC-3'

Oligo gspEEco RIrv

5'-AAGAATTCGTACACGGGCTACAGCC-3'

Oligo gspESrv

5'-AAGAGCTCCTTCACTGAGCATCG-3'

Oligo gspEfw

5'-GCAATTTAAGCAGCGCGAAGATG-3'

Oligo gspErv

5'-ATCTAGGGCACGGTATTCAAATGC-3'

PheSSNfw

5'-TTGTCGACATATGTCACATCTCGCAGAAC-3'

PheSXrv

5'-CCTCTAGAGAATTTCATAATCTATTCCTGCC-3'