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Endospore display platforms, products and methods

A technology for spores and nucleic acid molecules, applied in the field of endospore display platforms, products and methods, which can solve the problems that cannot be directly converted to the Bacillus family

Pending Publication Date: 2021-10-15
GINKGO BIOWORKS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high genetic diversity among bacteria, findings based on Bacillus subtilis studies often cannot be directly translated to family internal and external members of the Bacillus family

Method used

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  • Endospore display platforms, products and methods
  • Endospore display platforms, products and methods
  • Endospore display platforms, products and methods

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0205] Example 1: General protocol for the preparation of recombinant Brevibacillus, Lysinibacillus or Chlorobacillus endospores

[0206] To generate fusion constructs, polynucleotides encoding one or more heterologous proteins can be fused to polynucleotides encoding amino acids of any of the N-terminal targeting sequences disclosed herein, e.g., encoding Table 1 or Figure 1-3 (for Brevibacillus), Table 2 or Figure 4-5 (for Lysinibacillus) or Table 3 or Image 6 A polynucleotide of any amino acid sequence disclosed in (for Chlorobacillus genus), or a fragment or variant thereof which retains the exine targeting function when expressed in the respective bacterial genus. The fusion construct may be under the control of the native promoter of the disclosed N-terminal targeting sequence. Such constructs can be generated using splicing by overlap extension (SOE) techniques or Gibson assembly to generate linear amplicons. Protocols can also be modified at this stage to fuse the...

Embodiment 2

[0207] Example 2: Recombinant Brevibacillus, Lysinibacillus or Chlorobacillus endospores for delivery of fusion proteins involved in the production of plant growth promoting compounds to seeds, seedlings, plants or plant parts the use of

[0208] The enzymes responsible for the production of plant growth promoting compounds can be delivered to plants using the Brevibacillus, Lysinibacillus or Chlorobacillus endospore delivery systems disclosed herein. For example, butanediol dehydrogenase converts acetoin to 2,3-butanediol. 2,3-Butanediol is a compound that promotes plant growth. Brevibacillus, Lysinibacillus, or Chlorobacillus endospores expressing the enzyme can be applied as a seed treatment or seed coating, or delivered by drip or spray to the surrounding seeds, seedlings, plants or plant parts Area.

Embodiment 3

[0209] Example 3: Recombinant Brevibacillus, Lysinibacillus, or Chlorobacillus endospores for use on individual Brevibacillus, Lysinibacillus, or Chlorobacillus endospores Use of more than one fusion protein for delivery to seeds, seedlings, plants or plant parts

[0210] A single recombinant Brevibacillus, Lysinibacillus, or Chlorobacillus endospore can be used to display more than one heterologous fusion protein. This is achieved by constructing two (or more) separate fusion proteins. The coding sequence for each heterologous protein to be displayed on the surface of Brevibacillus, Lysinibacillus or Chlorobacillus endospores, respectively, was fused to an N-terminal targeting sequence under the control of its native promoter. Fusion protein constructs can be cloned into the same plasmid vector or into different plasmid vectors and introduced into Brevibacillus, Lysinibacillus or Chlorobacillus cells by electroporation. The resulting Brevibacillus, Lysinibacillus or Chlorob...

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PUM

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Abstract

Signal sequences useful for targeting proteins and peptides to the surface of endospores produced by Brevibacillus, Lysinibacillus, or Viridibacillus family members and methods of using the same are provided. The display of heterologous molecules, such as peptides, polypeptides and other recombinant constructs, on the exosporium of Brevibacillus, Lysinibacillus, or Viridibacillus family members, using particular N-terminal targeting sequences and derivatives of the same, and likewise are provided.

Description

[0001] Cross-references to related applications [0002] This application claims priority from U.S. Provisional Patent Application Nos. 62 / 768,077, 62 / 768,063, and 62 / 767,997, filed on November 15, 2018, the entire contents of each of which are incorporated herein by reference. [0003] Reference to an electronically submitted sequence listing [0004] Submit an official copy of the Sequence Listing electronically via EFS-Web as a Sequence Listing in ASCII format, file named "BCS189008WO_ST25", created on November 13, 2019, 169 kilobytes in size, and concurrent with this specification submit. The Sequence Listing contained in this ASCII format document is part of this specification and is incorporated herein by reference in its entirety. Technical field [0005] The present disclosure provides products useful for a variety of applications, such as delivering heterologous molecules of interest to plants. In particular, this disclosure describes endospore display methods and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N37/00C07K14/32
CPCC07K2319/02C07K14/195C12N15/74A01H3/00C12N15/62Y02A40/146A01N63/20C07K2319/50C12N15/625
Inventor B·L·戈洛姆F·A·菲卡拉R·萨拉姆扎德D·柯蒂斯K·蒂普顿D·艾利
Owner GINKGO BIOWORKS INC
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