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Table 2 Gateway recombination cloning of 34 LUCA and ECHH genes

From: Homologous high-throughput expression and purification of highly conserved E coli proteins

No.

Protein

Yield with His7-tag

Fusion with

Yield

157

aspartokinase I; homoserine dehydrogenase I

1

MBP

3

145

acetolactate synthase III small subunit

1

MBP

3

203

thiogalactoside acetyltransferase

1

MBP

2

200

4-hydroxy-2-ketovalerate aldolase

1

MBP

2

258

putative ATPase

1

MBP

3

204

anthranilate synthase component II

1

MBP

2

155

putative ATP-binding component of a transport system

1

MBP

3

201

flagellum-specific ATP synthase

1

MBP

2

259

fused enoyl-CoA hydratase and epimerase

1

MBP

2

156

FFh

1

MBP

2

146

30S ribosomal subunit protein S11

1

MBP

2

147

30S ribosomal subunit protein S13

1

MBP

3

148

50S ribosomal subunit protein L14

1

MBP

3

149

50S ribosomal subunit protein L22

1

MBP

3

151

50S ribosomal subunit protein L23

1

MBP

2

152

30S ribosomal subunit protein S12

1

MBP

2

150

50S ribosomal subunit protein L2

1

MBP

3

154

50S ribosomal subunit protein L1

1

MBP

3

153

biotin- [acetyl-CoA carboxylase] holoenzyme synthetase

1

MBP

3

159

RNA polymerase beta prime subunit

1

MBP

2

143

predicted acyltransferase with acyl-CoA N-acyltransferase

1

MBP

3

144

putative proteoglycan

1

MBP

2

158

endonuclease III

1

MBP

2

161

dihydroxyacid dehydratase

1

MBP

3

202

part of formate-dependent nitrite reductase complex

1

NusA

2

206

conserved protein, member of DEAD box family

1

NusA

2

198

50S ribosomal subunit protein L5

0

NusA

2

205

O-6-alkylguanine-DNA/cysteine-protein methyltransferase

0

NusA

1

199

acetylornithine delta-aminotransferase

0

NusA

1

197

inducible ATP-independent RNA helicase

1

GST

2

160

glutamate synthase large subunit

1

 

-

207

carbamoyl-phosphate synthase large subunit

1

 

-

208

ATP-binding transport protein (membrane protein)

0

 

-

209

Mg2+ transport ATPase, P-type 1 (membrane protein)

0

 

-

  1. Yields of purified proteins were analyzed by SDS-PAGE before recombination (proteins fused to His7-tag) and after recombination (each target protein fused to MBP-His7, GST-His7, or NusA-His6, respectively). The fusion partner that most efficiently improved protein yield is shown. With regard to yield, proteins were attributed to one of four categories: 3 (high), 2 (moderate), 1 (low), 0 (not purified).
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Microbial Cell Factories

ISSN: 1475-2859

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