Table 3 Bacteria, plasmids and oligonucleotide primers
Name | Characteristics | Reference or origin |
|---|---|---|
Janthinobacterium sp. 4239 | ||
Gram negative, chitosanolytic, no chitinolytic activity detected. | This study | |
E.coli | ||
DH10B | F' mcr A Δ(mrr-hsd RMS-mcr BC) Φ80lac ZΔM15 Δlac X74 deo R rec A1 end A1 ara D139 Δ(ara, leu)7697 gal U gal K λ- rps L (Streptomycin R) nup G | Invitrogen |
MGJ1042 | DH10B:pMGJ1042. Reference strain. | This study |
MGJ1061 | DH10B:pMGJ1061. Chromosomal library clone expressing Cho4239-1. | This study |
BMS172 | DH10B:pBMS172. Construction expressing Cho4239-1 fused with the OmpA signal peptide. | This study |
Plasmids | ||
pUC18 | lac UV5 promoter, Ampr, ColEI origin, 2.69 kb. | [40] |
pMGJ1042 | pMF1::OmpA-Sph I fragment. tetp/o promoter, Cmr, pro AB complementation and ColEI replication origin. | [36] |
pMGJ1061 | pUC18::4 kb chromosomal fragment encoding Cho4239-1. | This study |
pBMS172 | pMGJ1042::815 bp PCR fragment digested with Sph I/Bam HI and fused with OmpA signal peptide. | This study |
Oligonucleotide primers | ||
chodel4239-1.for | 5'-ACATGCATGCGGCGGCGGAGAACAGC-3' | Construction of pBMS172 |
cho4239-1.rev3 | 5'-CGCGGATCCGCGGCTGTTTAGCGGATCTGGT-3' | Construction of pBMS172 |
616V | 5'-AGAGTTTGATYMTGGCTCAG-3' | 16S RNA sequence |
630R | 5'-CAKAAAGGAGGTGATCC-3' | 16S RNA sequence |