Table 1.

Plasmids and oligonucleotides

Plasmids

pGME-T: Ampr, PCR cloning vector

pGA1: pGME-T Easy with 1,690 bp cheA1 gene amplified by PCR

pGA2: pGEM-T Easy with 2,521 bp cheA2 gene amplified by PCR

pGA1kan: pGA1 with 1.3kb flgB-kan replacing 393 bp HindIII/HindIII fragment

pGA2kan: pGA2 with 1.3kb flgB-kan replacing 1,326 bp HindIII/HindIII fragment

pGA2ery: pGA2 with 1.1 kb ermC replacing 1,326 bp HindIII/HindIII fragment

pQE-31: Expression vector

pQE-CheA1: cheA1 cloned into pQE-31 BamHI/PstI sites

DNA Primers:

5′-ATATTTAATCTCAATAGC-3′ (cheW2 reverse, primer extension)

5′-AAGTAATTTCTGAGATCG-3′ (cheA1, 5′ +, inactivation)

5′-AAAGTCTGATAACAGTTCGG-3′ (cheA1, 5′ +, inactivation)

5′-TAGTTATTGCATCTATGTC-3′ (cheA1, 3′ −, inactivation)

5′-ATGGAAATATTAGATTTGG-3′ (cheA2, 5′ +, inactivation)

5′-ATTTGGAAAATGAAGAGC-3′ (cheA2, 5′ +, inactivation)

5′-TTACCATTGCCAAGCGTAG-3′ (cheA2, 3′ −, inactivation)

5′-AAGCTTTAATACCCGAGCTTCAAG-3′ (flgB-kan, 5′ +, kan cassette)

5′-AAGCTTTCAAGTCAGCGTAATGCT-3′ (flgB-kan, 3′ −, kan cassette)

5′-AAGCTTAACACACTAGACTTATTTAC-3′ (ermC, 5′ +, ermC cassette)

5′-AAGCTTAAAAAATAGGCACACGAAAA-3′ (ermC, 3′ −, ermC cassette)

5′-ACGGATCCGGATAGTAGTGATGTTATTG-3′ (cheA1, 5′ +, expression)

5′-CTGCAGTATAAGTTTAGTTATTGCATC-3′ (cheA1, 3′ −, expression)

Italics indicate engineered restriction cut sites.