char = S;code = 83 char = u;code = 117 char = b;code = 98 char = c;code = 99 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = u;code = 117 char = l;code = 108 char = a;code = 97 char = r;code = 114 char = ;code = 32 char = L;code = 76 char = o;code = 111 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = i;code = 105 char = z;code = 122 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = G;code = 71 char = U;code = 85 char = S;code = 83 char = -;code = 45 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = G;code = 71 char = F;code = 70 char = P;code = 80 char = -;code = 45 char = T;code = 84 char = a;code = 97 char = g;code = 103 char = g;code = 103 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = P;code = 80 char = r;code = 114 char = o;code = 111 char = t;code = 116 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = s;code = 115 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = O;code = 79 char = n;code = 110 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = E;code = 69 char = p;code = 112 char = i;code = 105 char = d;code = 100 char = e;code = 101 char = r;code = 114 char = m;code = 109 char = a;code = 97 char = l;code = 108 char = ;code = 32 char = C;code = 67 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = s;code = 115 char = .;code = 46 result sql = Subcellular Localization of GUS- and GFP-Tagged Proteins in Onion Epidermal Cells.
Subcellular Localization of GUS- and GFP-Tagged Proteins in Onion Epidermal Cells.char = I;code = 73 char = N;code = 78 char = T;code = 84 char = R;code = 82 char = O;code = 79 char = D;code = 68 char = U;code = 85 char = C;code = 67 char = T;code = 84 char = I;code = 73 char = O;code = 79 char = N;code = 78 char = R;code = 82 char = e;code = 101 char = c;code = 99 char = o;code = 111 char = m;code = 109 char = b;code = 98 char = i;code = 105 char = n;code = 110 char = a;code = 97 char = n;code = 110 char = t;code = 116 char = ;code = 32 char = t;code = 116 char = a;code = 97 char = g;code = 103 char = s;code = 115 char = ;code = 32 char = (;code = 40 char = i;code = 105 char = .;code = 46 char = e;code = 101 char = .;code = 46 char = ,;code = 44 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = p;code = 112 char = o;code = 111 char = r;code = 114 char = t;code = 116 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = t;code = 116 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = s;code = 115 char = );code = 41 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = f;code = 102 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = ;code = 32 char = e;code = 101 char = x;code = 120 char = c;code = 99 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = e;code = 101 char = n;code = 110 char = t;code = 116 char = ;code = 32 char = a;code = 97 char = l;code = 108 char = t;code = 116 char = e;code = 101 char = r;code = 114 char = n;code = 110 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = v;code = 118 char = e;code = 101 char = ;code = 32 char = t;code = 116 char = o;code = 111 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = t;code = 116 char = i;code = 105 char = b;code = 98 char = o;code = 111 char = d;code = 100 char = i;code = 105 char = e;code = 101 char = s;code = 115 char = ;code = 32 char = f;code = 102 char = o;code = 111 char = r;code = 114 char = ;code = 32 char = d;code = 100 char = e;code = 101 char = t;code = 116 char = e;code = 101 char = r;code = 114 char = m;code = 109 char = i;code = 105 char = n;code = 110 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = s;code = 115 char = u;code = 117 char = b;code = 98 char = c;code = 99 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = u;code = 117 char = l;code = 108 char = a;code = 97 char = r;code = 114 char = ;code = 32 char = l;code = 108 char = o;code = 111 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = i;code = 105 char = z;code = 122 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = t;code = 116 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = s;code = 115 char = .;code = 46 char = ;code = 32 char = T;code = 84 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = m;code = 109 char = o;code = 111 char = s;code = 115 char = t;code = 116 char = ;code = 32 char = u;code = 117 char = s;code = 115 char = e;code = 101 char = r;code = 114 char = -;code = 45 char = f;code = 102 char = r;code = 114 char = i;code = 105 char = e;code = 101 char = n;code = 110 char = d;code = 100 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = t;code = 116 char = a;code = 97 char = g;code = 103 char = s;code = 115 char = ;code = 32 char = a;code = 97 char = r;code = 114 char = e;code = 101 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = &;code = 38 char = #;code = 35 char = 2;code = 50 char = 2;code = 50 char = 3;code = 51 char = ;;code = 59 char = -;code = 45 char = g;code = 103 char = l;code = 108 char = u;code = 117 char = c;code = 99 char = u;code = 117 char = r;code = 114 char = o;code = 111 char = n;code = 110 char = i;code = 105 char = d;code = 100 char = a;code = 97 char = s;code = 115 char = e;code = 101 char = ;code = 32 char = (;code = 40 char = G;code = 71 char = U;code = 85 char = S;code = 83 char = );code = 41 char = ;code = 32 char = r;code = 114 char = e;code = 101 char = p;code = 112 char = o;code = 111 char = r;code = 114 char = t;code = 116 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = e;code = 101 char = n;code = 110 char = z;code = 122 char = y;code = 121 char = m;code = 109 char = e;code = 101 char = ;code = 32 char = f;code = 102 char = r;code = 114 char = o;code = 111 char = m;code = 109 char = ;code = 32 char = E;code = 69 char = s;code = 115 char = c;code = 99 char = h;code = 104 char = e;code = 101 char = r;code = 114 char = i;code = 105 char = c;code = 99 char = h;code = 104 char = i;code = 105 char = a;code = 97 char = ;code = 32 char = c;code = 99 char = o;code = 111 char = l;code = 108 char = i;code = 105 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = f;code = 102 char = l;code = 108 char = u;code = 117 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = c;code = 99 char = e;code = 101 char = n;code = 110 char = t;code = 116 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = t;code = 116 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = s;code = 115 char = ;code = 32 char = d;code = 100 char = e;code = 101 char = r;code = 114 char = i;code = 105 char = v;code = 118 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = i;code = 105 char = m;code = 109 char = a;code = 97 char = r;code = 114 char = i;code = 105 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = f;code = 102 char = r;code = 114 char = o;code = 111 char = m;code = 109 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = g;code = 103 char = r;code = 114 char = e;code = 101 char = e;code = 101 char = n;code = 110 char = ;code = 32 char = f;code = 102 char = l;code = 108 char = u;code = 117 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = c;code = 99 char = e;code = 101 char = n;code = 110 char = t;code = 116 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = t;code = 116 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = (;code = 40 char = G;code = 71 char = F;code = 70 char = P;code = 80 char = );code = 41 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = ;code = 32 char = j;code = 106 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = y;code = 121 char = f;code = 102 char = i;code = 105 char = s;code = 115 char = h;code = 104 char = ;code = 32 char = A;code = 65 char = e;code = 101 char = q;code = 113 char = u;code = 117 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = a;code = 97 char = ;code = 32 char = v;code = 118 char = i;code = 105 char = c;code = 99 char = t;code = 116 char = o;code = 111 char = r;code = 114 char = i;code = 105 char = a;code = 97 char = .;code = 46 char = ;code = 32 char = G;code = 71 char = U;code = 85 char = S;code = 83 char = ;code = 32 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = u;code = 117 char = s;code = 115 char = e;code = 101 char = f;code = 102 char = u;code = 117 char = l;code = 108 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = i;code = 105 char = m;code = 109 char = a;code = 97 char = r;code = 114 char = i;code = 105 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = a;code = 97 char = s;code = 115 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = t;code = 116 char = a;code = 97 char = g;code = 103 char = ;code = 32 char = t;code = 116 char = o;code = 111 char = ;code = 32 char = a;code = 97 char = d;code = 100 char = d;code = 100 char = r;code = 114 char = e;code = 101 char = s;code = 115 char = s;code = 115 char = ;code = 32 char = n;code = 110 char = u;code = 117 char = c;code = 99 char = l;code = 108 char = e;code = 101 char = a;code = 97 char = r;code = 114 char = ;code = 32 char = l;code = 108 char = o;code = 111 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = i;code = 105 char = z;code = 122 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ,;code = 44 char = ;code = 32 char = w;code = 119 char = h;code = 104 char = e;code = 101 char = r;code = 114 char = e;code = 101 char = a;code = 97 char = s;code = 115 char = ;code = 32 char = G;code = 71 char = F;code = 70 char = P;code = 80 char = ;code = 32 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = m;code = 109 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = ;code = 32 char = v;code = 118 char = e;code = 101 char = r;code = 114 char = s;code = 115 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = l;code = 108 char = e;code = 101 char = .;code = 46 char = ;code = 32 char = M;code = 77 char = o;code = 111 char = r;code = 114 char = e;code = 101 char = o;code = 111 char = v;code = 118 char = e;code = 101 char = r;code = 114 char = ,;code = 44 char = ;code = 32 char = G;code = 71 char = F;code = 70 char = P;code = 80 char = ;code = 32 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = d;code = 100 char = e;code = 101 char = t;code = 116 char = e;code = 101 char = c;code = 99 char = t;code = 116 char = a;code = 97 char = b;code = 98 char = l;code = 108 char = e;code = 101 char = ;code = 32 char = d;code = 100 char = i;code = 105 char = r;code = 114 char = e;code = 101 char = c;code = 99 char = t;code = 116 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = l;code = 108 char = i;code = 105 char = v;code = 118 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = c;code = 99 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = s;code = 115 char = ,;code = 44 char = ;code = 32 char = w;code = 119 char = h;code = 104 char = e;code = 101 char = r;code = 114 char = e;code = 101 char = a;code = 97 char = s;code = 115 char = ;code = 32 char = G;code = 71 char = U;code = 85 char = S;code = 83 char = ;code = 32 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = o;code = 111 char = n;code = 110 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = d;code = 100 char = e;code = 101 char = t;code = 116 char = e;code = 101 char = c;code = 99 char = t;code = 116 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = i;code = 105 char = n;code = 110 char = d;code = 100 char = i;code = 105 char = r;code = 114 char = e;code = 101 char = c;code = 99 char = t;code = 116 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = b;code = 98 char = y;code = 121 char = ;code = 32 char = s;code = 115 char = t;code = 116 char = a;code = 97 char = i;code = 105 char = n;code = 110 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = f;code = 102 char = i;code = 105 char = x;code = 120 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = t;code = 116 char = i;code = 105 char = s;code = 115 char = s;code = 115 char = u;code = 117 char = e;code = 101 char = .;code = 46 char = ;code = 32 char = T;code = 84 char = h;code = 104 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = m;code = 109 char = a;code = 97 char = y;code = 121 char = ;code = 32 char = l;code = 108 char = e;code = 101 char = a;code = 97 char = d;code = 100 char = ;code = 32 char = t;code = 116 char = o;code = 111 char = ;code = 32 char = a;code = 97 char = r;code = 114 char = t;code = 116 char = i;code = 105 char = f;code = 102 char = a;code = 97 char = c;code = 99 char = t;code = 116 char = s;code = 115 char = ;code = 32 char = o;code = 111 char = r;code = 114 char = ;code = 32 char = i;code = 105 char = t;code = 116 char = ;code = 32 char = m;code = 109 char = a;code = 97 char = y;code = 121 char = ;code = 32 char = o;code = 111 char = b;code = 98 char = s;code = 115 char = c;code = 99 char = u;code = 117 char = r;code = 114 char = e;code = 101 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = b;code = 98 char = l;code = 108 char = e;code = 101 char = m;code = 109 char = s;code = 115 char = ;code = 32 char = w;code = 119 char = i;code = 105 char = t;code = 116 char = h;code = 104 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = t;code = 116 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = s;code = 115 char = o;code = 111 char = l;code = 108 char = u;code = 117 char = b;code = 98 char = i;code = 105 char = l;code = 108 char = i;code = 105 char = t;code = 116 char = y;code = 121 char = .;code = 46 char = ;code = 32 char = I;code = 73 char = n;code = 110 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = t;code = 116 char = o;code = 111 char = c;code = 99 char = o;code = 111 char = l;code = 108 char = ,;code = 44 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = t;code = 116 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = l;code = 108 char = o;code = 111 char = c;code = 99 char = a;code = 97 char = l;code = 108 char = i;code = 105 char = z;code = 122 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = r;code = 114 char = o;code = 111 char = u;code = 117 char = t;code = 116 char = i;code = 105 char = n;code = 110 char = e;code = 101 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = a;code = 97 char = s;code = 115 char = s;code = 115 char = a;code = 97 char = y;code = 121 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = a;code = 97 char = f;code = 102 char = t;code = 116 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = p;code = 112 char = a;code = 97 char = r;code = 114 char = t;code = 116 char = i;code = 105 char = c;code = 99 char = l;code = 108 char = e;code = 101 char = -;code = 45 char = m;code = 109 char = e;code = 101 char = d;code = 100 char = i;code = 105 char = a;code = 97 char = t;code = 116 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = t;code = 116 char = r;code = 114 char = a;code = 97 char = n;code = 110 char = s;code = 115 char = i;code = 105 char = e;code = 101 char = n;code = 110 char = t;code = 116 char = ;code = 32 char = t;code = 116 char = r;code = 114 char = a;code = 97 char = n;code = 110 char = s;code = 115 char = f;code = 102 char = o;code = 111 char = r;code = 114 char = m;code = 109 char = a;code = 97 char = t;code = 116 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = o;code = 111 char = f;code = 102 char = ;code = 32 char = o;code = 111 char = n;code = 110 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = e;code = 101 char = p;code = 112 char = i;code = 105 char = d;code = 100 char = e;code = 101 char = r;code = 114 char = m;code = 109 char = a;code = 97 char = l;code = 108 char = ;code = 32 char = c;code = 99 char = e;code = 101 char = l;code = 108 char = l;code = 108 char = s;code = 115 char = .;code = 46 char = ;code = 32 char = W;code = 87 char = i;code = 105 char = t;code = 116 char = h;code = 104 char = ;code = 32 char = t;code = 116 char = h;code = 104 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = m;code = 109 char = e;code = 101 char = t;code = 116 char = h;code = 104 char = o;code = 111 char = d;code = 100 char = ;code = 32 char = i;code = 105 char = t;code = 116 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = n;code = 110 char = ;code = 32 char = b;code = 98 char = e;code = 101 char = ;code = 32 char = d;code = 100 char = e;code = 101 char = t;code = 116 char = e;code = 101 char = r;code = 114 char = m;code = 109 char = i;code = 105 char = n;code = 110 char = e;code = 101 char = d;code = 100 char = ;code = 32 char = r;code = 114 char = a;code = 97 char = p;code = 112 char = i;code = 105 char = d;code = 100 char = l;code = 108 char = y;code = 121 char = ;code = 32 char = w;code = 119 char = h;code = 104 char = e;code = 101 char = t;code = 116 char = h;code = 104 char = e;code = 101 char = r;code = 114 char = ;code = 32 char = a;code = 97 char = ;code = 32 char = g;code = 103 char = i;code = 105 char = v;code = 118 char = e;code = 101 char = n;code = 110 char = ;code = 32 char = f;code = 102 char = u;code = 117 char = s;code = 115 char = i;code = 105 char = o;code = 111 char = n;code = 110 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = o;code = 111 char = t;code = 116 char = e;code = 101 char = i;code = 105 char = n;code = 110 char = ;code = 32 char = i;code = 105 char = s;code = 115 char = ;code = 32 char = a;code = 97 char = c;code = 99 char = t;code = 116 char = i;code = 105 char = v;code = 118 char = e;code = 101 char = ,;code = 44 char = ;code = 32 char = a;code = 97 char = n;code = 110 char = d;code = 100 char = ;code = 32 char = p;code = 112 char = r;code = 114 char = e;code = 101 char = l;code = 108 char = i;code = 105 char = m;code = 109 char = i;code = 105 char = n;code = 110 char = a;code = 97 char = r;code = 114 char = y;code = 121 char = ;code = 32 char = t;code = 116 char = a;code = 97 char = r;code = 114 char = g;code = 103 char = e;code = 101 char = t;code = 116 char = i;code = 105 char = n;code = 110 char = g;code = 103 char = ;code = 32 char = d;code = 100 char = a;code = 97 char = t;code = 116 char = a;code = 97 char = ;code = 32 char = c;code = 99 char = a;code = 97 char = n;code = 110 char = ;code = 32 char = b;code = 98 char = e;code = 101 char = ;code = 32 char = o;code = 111 char = b;code = 98 char = t;code = 116 char = a;code = 97 char = i;code = 105 char = n;code = 110 char = e;code = 101 char = d;code = 100 char = .;code = 46 char = <;code = 60 char = b;code = 98 char = r;code = 114 char = /;code = 47 char = >;code = 62 result sql = INTRODUCTIONRecombinant tags (i.e., reporter proteins) offer an excellent alternative to antibodies for determining the subcellular localization of proteins. The most user-friendly tags are the ß-glucuronidase (GUS) reporter enzyme from Escherichia coli and fluorescent proteins derived primarily from the green fluorescent protein (GFP) of the jellyfish Aequorea victoria. GUS is useful primarily as a tag to address nuclear localization, whereas GFP is more versatile. Moreover, GFP is detectable directly in living cells, whereas GUS is only detected indirectly by staining of fixed tissue. This may lead to artifacts or it may obscure problems with protein solubility. In this protocol, protein localization is routinely assayed after particle-mediated transient transformation of onion epidermal cells. With this method it can be determined rapidly whether a given fusion protein is active, and preliminary targeting data can be obtained.
Publication: CSH Protoc 2007 ;2007:pdb.prot4689. :doi: 10.1101/pdb.prot4689. :pii: 2007/2/pdb.prot4689.
Publication Date: 2007
Study Author(s): von Arnim, Albrecht;
Institution:
Shortcut link to this study:
INTRODUCTIONRecombinant tags (i.e., reporter proteins) offer an excellent alternative to antibodies for determining the subcellular localization of proteins. The most user-friendly tags are the ß-glucuronidase (GUS) reporter enzyme from Escherichia coli and fluorescent proteins derived primarily from the green fluorescent protein (GFP) of the jellyfish Aequorea victoria. GUS is useful primarily as a tag to address nuclear localization, whereas GFP is more versatile. Moreover, GFP is detectable directly in living cells, whereas GUS is only detected indirectly by staining of fixed tissue. This may lead to artifacts or it may obscure problems with protein solubility. In this protocol, protein localization is routinely assayed after particle-mediated transient transformation of onion epidermal cells. With this method it can be determined rapidly whether a given fusion protein is active, and preliminary targeting data can be obtained.
PMID: 21357024