TABLE 1.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant genotype or characteristicsSource or referencea
Strains
    E. coli DH5αF φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17 phoA supE44 thi-1 gyrA96 relA1Amersham Pharmacia Biotech, Piscataway, N.J.
    E. coli BL21F ompT hsdSB(rBmB) gal dcmAmersham Pharmacia Biotech, Piscataway, N.J.
    Sphingobium indicum B90AYellow colonies, produces brown pigment, fast degrader of HCH isomersN. Sethunathan, CRRI, Cuttack, India (41)
    Sphingobium japonicum UT26Yellow colonies, degrades HCH isomers, albeit slowlyY. Nagata, University of Tokyo, Tokyo, Japan (44)
    Sphingobium francense Sp+Yellow colonies, degrades HCH isomers, albeit slowlyTim Vogel, University of Lyon, Lyon, France (6)
Plasmids
    pUC182.7 kb; Ampr; multiple cloning site internal to lacZ geneFermentas Inc.
    pUC4K3.9 kb; Ampr Kanr pBR322 ori; restriction site mobilizing elementAmersham Pharmacia Biotech, Piscataway, N.J.
    pUCISpUC18 containing 1-kb IS6100 element (amplified from pLINA57)This study
    pUCIS1KpUCIS containing 1.2-kb Kanr fragment from pUC4KThis study
    pUCIS1KBpUCIS1K containing linB of S. indicum B90A cloned at KpnI siteThis study
    pGEX-5X-34.9 kb; GST fusion vector with tac promoter, lacIq, and factor Xa protease recognition siteAmersham Pharmacia Biotech, Piscataway, N.J.
    pLINB35pWE15 carrying DNA fragment of B90A containing linB and IS61008
    pLINA57pWE15 carrying 41-kb DNA fragment of B90A containing linA1, linC, linX, and IS61008
    pLINSBpUC18 containing HindIII-digested 2.16-kb fragment of S. francense Sp+ containing linB ORF and IS6100This study
    pLINEBBlinB ORF of S. indicum B90A cloned into pGEX-5X-3 at BamHI and XhoI sitesThis study
    pLINEUBlinB ORF of S. japonicum UT26 cloned into pGEX-5X-3 at BamHI and XhoI sitesThis study
    pLINESBlinB ORF of S. francense Sp+ cloned into pGEX-5X-3 at EcoRI and XhoI sitesThis study
Mutants
    Sp+mtSp+ mutant lacking linA and linB, degrades none of the four HCH isomers8
    Sp+mtC1-9Sp+mt complemented with linB of B90A, degrades only β- and δ-HCH isomersThis study
  • a CRRI, Central Rice Research Institute.