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- あゆみ たなせ
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34 (Ralstonia, Hydrogenovibrio ) (Synechococcus, Spirulina ) (Thiobacillus ) (Rhodobacter, Rhodospirillum ) (Nitrosomonas ) (Chromatium ) (Nitrobacter ) ( Methanobacterium ) Acetyl-CoA (Clostridium ) 3-HP (Chloroflexus ) (3-hydroxypropionate cycle) (Acidianus, Sulfolobus ) TCA (Hydrogenobacter ) (Chlorobium ) (RTCA cycle) (Desulfobacter ) 21
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68 Image: George Retseck 55
69 Subsurface lithoautotrophic microbial ecosystems exist in the pores between interlocking mineral grains of many igneous rocks. Autotrophic microbes (green) derive nutrients and energy from inorganic chemicals in their surroundings, and many other microbes (red), in turn, feed on organics created 56
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114 modified seqence 101
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117 104
118 Clostridium acetobutylicum OXO seqence 105
119 106
120 107
121 TCA 108
122 109
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124 111
125 112
126 1 T. kodakaraensis KOD1 113
127 114
128 115
129 KOD1 116
130 KOD1ORF map 117
131 118
132 119
133 KU25 Uracil 120
134 KW4 pyrf 121
135 KW4 Southern blot 122
136 123
137 124
138 125
139 126
140 Rubisco 127
141 Tk-Rubisco 128
142 129
143 28 Tk-Rubisco Tk-Rubisco 130
144 131
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146 133
147 134
148 135
149 J Biol Chem Aug 30;277(35):
150 FOR 137
151 138
152 FOR 139
153 140
154 141
155 Tk-FOR 142
156 143
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158
159 146
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163 150
164 HD-1 151
165 16SrRNAProteobacteria 152
166 153
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170 157
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174 161
175 pmg105 pmg103 pmg
176 PEPCK 163
177 PEPCK 164
178 165
179 166
180 167
181 pm3rbcwt A TN TC PCR B RP TC PCR 168
182 169
183 pm3rbcwt R. palustris A TN TC PCR B RP TC PCR 170
184 171 Tk-Rbc Western blot 30 S HS Tk-Rbc SDS-PAGE S: HS:8515 HP: Rb: Tk-Rbc 1, 2 Tk-Rbc
185 R. palustris Triple mutant pm3rbcwt 172
186 PCR 10 PCR pm3rbcwt PEPCKpromoter-Tk-rbc DNA Colony No
187 PCR PCR Colony No. 1~6 Colony No.7 pm3rbcwt Colony No.8 Triple Mutant RP, TC, TN, 950r : PEPCK promoter, Tk-RbcC, Tk-Rbc N, Tk-Rbc 1kb 174
188 175
189 176
190 PCR pm3rbcwt 1: 2~5: 6~12: YE-acid N: R. palustris Triple mutant (Negative control)p: pm3rbcwt (Positive control)m: λ/hindiii Tk-Rbc Western blot 6~12: 35 6~12 P: Positive 177
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198 185
199 pem7/zeo MCS: 186
200 PCR pem7/zeo primer 2 3primer 1 3 PCR SalI site 187
201 pm3rbcwt EM7 promoter PEPCK promoter 188
202 PCR pm3ezrbcwt, pm3rzrbcwt PCR 189
203 R. palustris Triple mutant 190
204 Tk-Rbc Western blot Tk-Rbc (positive control)triple mutant pm3ezrbcwt YE-acid YE-acid 191
205 192
206 in vitro 193
207 194
208 Tk-Rubisco R. palustris Rubisco Double mutant DNA Rubisco 195
209 DNA Rubisco formi-2 1~5 DNA PCR DNA 400ng/l 1l DNA 1, 200ng. 2, 100ng. 3, 50ng. 4, 25ng. 5, 12.5ng Triple mutant DNA 196
210 Rubisco 197
211 R. palustris form I-2 Rubisco 198
212 pem7zeo/tk-rbc BglII, SalI Tk-rbc DNA 199
213 pwdzrl2tk-rbc 200
214 PCR Insert Check BamHI 201
215 202
216 203
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218 205
219 KOD1 Rubisco Rubisco Rubisco Rhodopseudomonas palustris Rubisco Rubisco Rubisco R. palustris Tk-rbc Rubisco Tk-rbc Tk-rbc Tk-rbc Rubisco R. palustris Tk-Rbc TypeIII Rubisco Rubisco Tk-Rubisco Tk-rbc ORF error prone PCR in vitro Tk-Rubisco R. palustris DNA Tk-rbc in vitro, in vivo 206
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231 RITE FAX 90
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12 ( 1) 70 1 ( 2) 2 4 4 45 38 3 3 3 6000 500 500-50 +50
9:11-38 2009 11 2008 12 26 3 46 12 ( 1) 70 1 ( 2) 2 4 4 45 38 3 3 3 6000 500 500-50 +50 13 38 5 19 5 19 Archaea/Eucarya 9 11 12 300 300 12 31 7 6 3 14 6 ( 8) 1973 1977 16S ribosomal RNA 2 90 80 8 ( 9)
20 15 14.6 15.3 14.9 15.7 16.0 15.7 13.4 14.5 13.7 14.2 10 10 13 16 19 22 1 70,000 60,000 50,000 40,000 30,000 20,000 10,000 0 2,500 59,862 56,384 2,000 42,662 44,211 40,639 37,323 1,500 33,408 34,472
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