Table I.

Summary of the E. coli strains, plasmids, and oligonucleotides used in this study

MaterialPropertyReference or Source
E. colistrain
 UTH903 λhisI903 rpsL145(strR) malT1(lR) malT1(lR) xy1A5 mtl-1 CGSC-a
 JM109 recA1 endA1 gyrA96 thi hsdR17 supE44 relA1 Δ(lac-proAB)/F′ Sambrook et al. (1989)
(traD36 proAB lacIqlacZΔM15)
 XLI-Blue hsdR17 endA1 supE44 thi recA1 relA1 lac/F′ (proAB lacIq lacZΔM15: :Tn10) Stratagene
gyrA96 relA1 lac[F′ proAB lacIq ZΔM15 Tn10]
 BL21 FompT[lon] hsdS Pharmacia
Plasmid
 pKF323 pBluescript bearing a 1.1-kb fragment for the At-IE cDNA (pAt-IE) This study
 pKF347 pMAL-c2 bearing a full-length ORF This study
 pKF360 pBluscript bearing a 5.5-kb fragment for the At-IE geneThis study
 pKF362pMAL-c2 bearing an N-terminal segment without its putative chloroplast transit peptideThis study
 pKF363pMAL-c2 bearing a C-terminal segmentThis study
 pKF371pMAL-c2 bearing an N-terminal segmentThis study
 pKF372pMAL-c2 bearing an ORF without its putative chloroplast transit peptide regionThis study
Oligonucleotide
 PRI28-b 5′-CGGAATTCGGATCCATGGCGGTATCGTACA-3′This study
 PRI30-c 5′-CGCTCGAGCAAAGTTGCACAATGAACAAC-3′This study
 PRI44-d 5′-GCGGATCCGTAGATAACTTGTTGGACCG-3′This study
 PRI45-e 5′-CGCTCGAGCTACTCATCATTGTTTAATTGA-3′This study
 PRI46-f 5′-GCGGATCCGCTTCAGGAAACAAGCTAGC-3′This study
  • F0-a E. coli Genetic Stock Center, Yale University, New Haven, CT.

  • F0-b Corresponding to the bases +1 to +16 of the At-IE gene on pKF360.

  • F0-c Corresponding to the bases +236 to +2346 of the At-IE gene on pKF360.

  • F0-d Corresponding to the bases +1343 to +1362 of the At-IE gene on pKF360.

  • F0-e Corresponding to the bases +1760 to +1742 of the At-IE gene on pKF360.

  • F0-f Corresponding to the bases +1855 to +1874 of the At-IE gene on pKF360.