An encoded N-terminal extension results in low levels of heterologous protein production in Escherichia coli

Table 1 Oligonucleotide primers used in this study^a

Oligonucleotide name	Sequence (5'->3')^b	Target ^c
BamHI nilC	GGATCC GACGATGCCTTAATGCGACAG	nilC
BamHI tdk	GGATCC CTTAGTGATTTTATACG	3' of tdk
clpP tetKWfor^d	ATCGGTACAGCAGGTTTTTTCAATTTTATCCAGGAGACGG CAAGAGGGTCATTATATTTCG	5' of E. coli clpP/Tn10 (tet^r)
clpP tetKWrev^d	GCCGCCCTGGATAAGTATAGCGGCACAGTTGCGCCTCTGG GACTCGACATCTTGGTTACCG	3' of E. coli clpP/ Tn10 (tet^r)
Ecrecaminfor	GAAAGCGGAAATCGAAGGCG	E. coli recA
Ecrecaminrev	CATCACACCAATTTTCATACGG	E. coli recA
EctdkQPfor	TTTGGTGCCGGGAAAGTC	E. coli tdk
EctdkQPrev	CTTGTTGTCTGGTTAAAAACTGG	E. coli tdk
NcoI MAGPNilC	CCATGG CCGGGCCAGCTAGAGGAGGGGGTTCTCACC	nilC
NcoI MDAPNilC	CCATGG ACGCGCCAGCTAGAGGAGGGGGTTCTCACC	nilC
NcoI MDGANilC	CCATGG ACGGGGCAGCTAGAGGAGGGGGTTCTCACC	nilC
NcoI MDGPNilC	CCATGG ACGGGCCAGCTAGAGGAGGGGGTTCTCACC	nilC
NcoI NilC	CCATGG CTAGAGGAGGGGGTTCTCACC	nilC
tdkprext2	AATAAAAATAAAGCTGAGCC	tdk
XntdkQPfor2	CTATCCGCTGATGCTTTGTTG	tdk
XntdkQPrev2	TACAATTTCACAAAGCTGCTC	tdk

^aAdditional oligonucleotides used in this study are listed in Table 2
^bUnderlined nucleotides indicate engineered restriction sites used in cloning
^cOligonucleotide primers are specific for X. nematophila unless noted otherwise; Integrated DNA Technologies (Coralville, IA) synthesized the oligonucleotides used in this study.
^dItalicized nucleotides indicate clpP-region sequence

ISSN: 1475-2859