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Dual transit peptides for targeting polypeptides

A technology for transporting peptides and objects, which is applied in the field of plant molecular biology and can solve problems such as organelle destruction

Pending Publication Date: 2019-01-04
BASF AGRO BV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Even though there may be some proteins that are theoretically shared by both organelles, most proteins have specific functions in the organelles they target, so that mistargeting may be counter-selected as unnecessary loss of valuable proteins, or even Destructive to organelles (Peeters and Small, Biochimica et Biophysica Acta 1541 (2001), pp. 54-63)

Method used

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  • Dual transit peptides for targeting polypeptides
  • Dual transit peptides for targeting polypeptides
  • Dual transit peptides for targeting polypeptides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0444] Example 1: Cloning of expression cassettes

[0445] 编码包含SEQ ID NO:112,113,114,115,116,117,118,119,120,121,122,123,124,125,126,127,128,129,130,131,132,133,134,136,592,593,594,595,596,597,598,599,600,601,602,603,604,605,606,607,608,609,610,611,612,613,614,615,616,617,618,619,620,621,622,623或624的多肽的全部核酸编码序列均由Geneart(Geneart AG,Regensburg,Germany)合成和克隆。 Rationally designed mutants were synthesized by Geneart. The random PPO gene library was synthesized by Geneart. Plasmids were isolated from E. coli TOP10 by performing a plasmid miniprep and confirmed by DNA sequencing.

Embodiment 2

[0446] Example 2: Engineering construction of herbicide-tolerant plants containing the expression cassette of the present invention

[0447]Herbicide tolerant soybeans (Glycine max), corn (Zea mays), and canola (Brassica napus) or turnip (Brassica Rapa var.) or Brassica campestris L.) plants. For transformation of soybean or Arabidopsis, the expression cassette / construct comprising the chimeric nucleic acid molecule is cloned into a binary vector using standard cloning techniques as described by Sambrook et al. (Molecular cloning (2001) Cold Spring Harbor Laboratory Press) 中,所述嵌合核酸分子编码包含SEQ ID NO:112,113,114,115,116,117,118,119,120,121,122,123,124,125,126,127,128,129,130,131,132,133,134,136,592,593,594,595,596,597,598,599,600,601,602,603,604,605,606,607,608,609,610,611,612,613,614,615,616,617,618,619,620,621,622,623或624的多肽,所述二元载体包含在泛素启动子(PcUbi)和胭脂碱合酶终止子(NOS)序列之间的抗性 Marker gene cassette (AHAS) and chimeric polypeptide sequence (labeled GOI). For maize transformation, the wild-t...

Embodiment 3

[0449] Example 3. Tissue Culture Conditions

[0450] An in vitro tissue culture mutagenesis assay has been developed to isolate and characterize the effects of protoporphyrinogen oxidase-inhibiting herbicides (trifludimoxazine, saflufenacil, flumioxazin, butafenacil ), acifluorfen, lactofen, bifenox, sulfentrazone, and photosynthesis inhibitor diuron (as a negative control) ) tolerant plant tissues (such as corn and rice tissues). The assay uses somaclonal variation found in in vitro tissue culture. Spontaneous mutations derived from somaclonal variation can be enhanced by chemical mutagenesis followed by stepwise selection on increasing concentrations of herbicides.

[0451] The present invention provides tissue culture conditions for promoting the growth of regenerable fragile embryogenic maize or rice callus. Calli were initiated from 4 different maize or rice cultivars including maize (Zea mays) and Japonica (Taipei 309, Nipponbare, Koshihikari) and Indica (Indica 1 ) v...

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Abstract

The present invention refers to a recombinant chimeric nucleic acid molecule comprising a nucleic acid sequence encoding a dual transit peptide operably linked to a heterologous nucleic acid sequenceencoding a polypeptide of interest which, when overexpressed in a plant, confers herbicide tolerance to said plant.

Description

Field of invention [0001] The present invention relates to the field of plant molecular biology, and more specifically to targeting polypeptides of interest to chloroplasts and / or mitochondria through the use of novel dual transport peptides. Background technique [0002] Plant cells contain two organelles originally derived from endosymbiotic bacteria: mitochondria and plastids. Due to their endosymbiotic origin, these organelles contain their own DNA, but only a few genes are actually encoded by these genomes. Many other genes that were originally present have been transferred into the host's nuclear genome, and the products of their expression are targeted back to the corresponding organelles. Although targeting of proteins to mitochondria and chloroplasts is often highly specific, a growing number of examples have been found in which the same proteins are imported into both organelles (Peeters and Small, Biochimica et Biophysica Acta 1541 (2001), p. 54 -page 63). Thes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82
CPCC12N15/8221C12N15/8274A01H1/00A01H5/00C12N15/8234
Inventor R·阿朋特S·特雷施T·塞萨尔J·M·保利克
Owner BASF AGRO BV
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