Purification of recombinant AtGA3OX4 shown by SDS-PAGE on a 8–15% gradient gel with Coomassie Blue staining

<p><b>Copyright information:</b></p><p>Taken from "Purification and kinetic studies of recombinant gibberellin dioxygenases"</p><p>BMC Plant Biology 2005;5():19-19.</p><p>Published online 25 Sep 2005</p><p>PMCID:PMC1262730.</p><p>Copyright © 2005 Lester et al; licensee BioMed Central Ltd.</p>(A) Lanes: 1, soluble cell fraction; 2, TRX-AtGA3OX4 after IMAC chromatography; 3, TRX-AtGA3OX4 after thrombin digest; 4, concentrated TRX-AtGA3OX4 (10 mg/ mL); 5, Bio Rad molecular weight standards, (from top) 115, 93, 49.8, 35.8, 29.2, 21.3 and 6.4 kDa.(B) Lanes: 1, TRX-AtGA3OX4 after thrombin digest; 2, AtGA3OX4 (1.5 mg/mL, maximal concentration achieved) after gel filtration on Superdex 75; 3, TRX-AtGA3OX4 (10 mg/mL) after gel filtration on Superdex 200. The molecular weights expected for TRX-AtGA3OX4, AtGA3OX4 and TRX are 54.1, 39.1 and 15 kDa, respectively. The arrows indicate the position of AtGA3OX4

Frequency of typical mutations observed among the 2,610 M₂ individuals screened.

<p>Frequency of typical mutations observed among the 2,610 M₂ individuals screened.</p

Mutation detection and estimation of mutation frequency in three candidate genes of <i>Ppd-D1</i>, <i>Rubisco activase A</i> and <i>Rubisco activase B</i> by TILLING analysis.

a<p>, mutation detected by non-denaturing polyacrylamide gels stained with silver;</p>b<p>, mutation detected by non-denaturing polyacrylamide gels stained with ethidium bromide;</p>c<p>, mutation detected by agarose gels stained with ethidium bromide.</p>*<p>For calculation of the mutation frequency, 100 bp sequences from each end were removed due to the base ambiguity.</p

Digested bands detected with non-denaturing polyacrylamide gels stained with either silver (a) or ethidium bromide (b) and agarose gels stained with ethidium bromide (c).

<p>Putative mutations in the pools (1, 2, 3, 4) are identified by the presence of two bands (indicated by white arrows), with sizes adding up to the full length PCR product. (a). Non-denaturing polyacrylamide gel stained with silver; (b). Non-denaturing polyacrylamide gel stained with ethidium bromide; (c). Agarose gels stained with ethidium bromide.</p

The mutation density and germination rate in published TILLING populations of different species under different EMS dosage treatments.

*<p>in M₃ plant.</p

Primer sequences of the 6 primers used and the mutation frequency detected.

<p>Primer sequences of the 6 primers used and the mutation frequency detected.</p

Analysis of mutations identified in the <i>Ppd-D1</i> gene.

*<p>Het, heterozygote; Hom, homozygote.</p><p>PSSM or SIFT scores of mutation lines with star (*) are predicted to be damaging to protein function. Mutation with PSSM score larger than 10 indicates that the mutation is more likely to have a damaging effect on the protein function. Mutation with SIFT score less than 0.05 is predicted to be deleterious.</p

Type and distribution of induced mutations discovered in <i>Ppd-D1</i> amplicon.

<p>Type and distribution of induced mutations discovered in <i>Ppd-D1</i> amplicon.</p

Mutant phenotypes observed in the M₂ and M₃ wheat plants.

<p>Mutant phenotypes: (a), (i), (j) dwarf and semi-dwarf; (b) single tiller; (c) coleoptile color; (d) seed size; (e), (m) albinism; (f) spike morphology; (g)(h) erect leaf; (k) narrow leaf; (l) strange leaf morphology; (n) disease sensitive; (o) large spikes with short awns; (p) yellow spots on leaves.</p

An example of RAPD banding pattern obtained by primer R1 and primer IT31.

<p>W: wild type; m: mutagenised lines; M: molecular weight standards DL2000.</p