| 主权项 |
1. A tomato plant comprising one or more cells in which an endogenous mitochondrial malate dehydrogenase (mMDH) gene is homozygously mutated by transforming the one or more cells with a vector encoding a pair of zinc finger nucleases, each zinc finger nuclease comprising a zinc finger protein having five or six zinc finger domains ordered finger 1 to finger 5 (F1 to F5) or finger 1 to finger 6 (F1 or F6), each zinc finger domain comprising a recognition helix region, wherein the zinc finger proteins comprise the following recognition helix regions:
(i) F1: RSDDLSE (SEQ ID NO:11);
F2; TNSNRKR (SEQ ID NO:12);F3: RSDHLST (SEQ ID NO:13);F4; TNSNRIT (SEQ ID NO:14);F5: RREDLIT (SEQ ID NO:15); andF6: TSSNLSR (SEQ ID NO:16), wherein the zinc finger protein binds to a target site as shown in SEQ ID NO:3; or (ii) F1: QSSDLSR (SEQ ID NO:17);
F2: TSGNLTR (SEQ ID NO:18);F3: RSDYLSK (SEQ ID NO:19);F4: TSSVRTT (SEQ ID NO:20);F5: TSGNLTR (SEQ ID NO:18); andF6: QRSHLSD (SEQ ID NO:22), wherein the zinc finger protein binds to a target site as shown in SEQ ID NO:4; or (iii) F1: RSDTLSV (SEQ ID NO:23);
F2: DNSTRIK (SEQ ID NO:24);F3; RSDHLSE (SEQ ID NO:25);F4: TSGSLTR (SEQ ID NO:26);F5: RSDALSR (SEQ ID NO:27); andF6: TSGNLTR (SEQ ID NO:18), wherein the zinc finger protein binds to a target site as shown in SEQ ID NO:5; or (iv) F1: RSDNLAR (SEQ ID NO:29);
F2: QRGNRNT (SEQ ID NO:30);F3: DSSDRKK (SEQ ID NO:31);F4: DRSNLSR (SEQ ID NO:32); andF5: LRHHLTR (SEQ ID NO:33), wherein the zinc finger protein binds to a target site as shown in SEQ ID NO:6; or (v) F1: DRSNLSR (SEQ ID NO:32);
F2: LRQNLIM (SEQ ID NO:35);F3: RSDALSE (SEQ ID NO:36);F4: RSSTRKT (SEQ ID NO:37);F5: DRSALSR (SEQ ID NO:38); andF6: RSDALAR (SEQ ID NO:39), wherein the zinc finger protein binds to a target site as shown in SEQ ID NO:7; or (vi) F1: QSGNLAR (SEQ ID NO:40);
F2: NRYDLHK (SEQ ID NO:41);F3: DRSNLSR (SEQ ID NO:32);F4: LRFARDA (SEQ ID NO:43);F5: RSDNLAR (SEQ ID NO:29); andF6: RSDHLTQ (SEQ ID NO:45), wherein the zinc finger protein binds to a targgt site as shown in SEQ ID NO:8; or (vii) F1: DRSDLSR (SEQ ID NO:46);
F2: QAGNLKK (SEQ ID NO:47);F3: QSGSLTR (SEQ ID NO:48);F4: RSDNLRE (SEQ ID NO:49); andF5: DSSDRKK (SEQ ID NO:31), wherein the zinc finger protein binds to a target site as shown in SEQ ID NO:9; or (viii) F1: DRSNLSR (SEQ ID NO:32);
F2: LKQHLTR (SEQ ID NO:52);F3: QSSDLSR (SEQ ID NO:17);F4: QSGNLAR (SEQ ID NO:40);F5: RSDHLSQ (SEQ ID NO:55); andF6: QNAHRIT (SEQ ID NO:56), wherein the zinc finger protein binds to a target site as shown in SEQ ID NO:3 such that activity of an expressed mMDH protein is reduced and wherein crop yield from the tomato plant is increased. |
