Novel synthetic genes for plant gums

a technology of plant gums and synthetic genes, applied in the field of plant gum synthetic genes, can solve the problems of affecting the growth rate of plant gums, so as to achieve the effect of maximizing the number of cells receiving

Inactive Publication Date: 2004-01-15
OHIO UNIV TECH TRANSFER OFFICE TECH & ENTERPRISE BUILDING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

0107] Other particle bombardment methods are also available for the introduction of heterologous nucleic acid sequences into plant cells. Generally, these methods involve depositing the nucleic acid sequence of interest upon the surface of small, dense particles of a material such as gold, platinum, or tungsten. The coated particles are themselves then coated onto either a rigid surface, suc...

Problems solved by technology

It is a tedious process involving piercing and stripping the bark of the trees, then returning later to gather the dried tear drop shaped, spherical balls that form.
However, this production depends on the environmental and political stability of the region producing the gum.
Again in 1...

Method used

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  • Novel synthetic genes for plant gums
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  • Novel synthetic genes for plant gums

Examples

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example 2

Construction of Synthethic HRGP Gene Cassettes

[0121] Synthetic gene cassettes encoding contiguous and noncontiguous Hyp modules are constructed using partially overlapping sets consisting of oligonucleotide pairs, "internal repeat pairs" and "external 3'- and 5'-linker pairs" respectively, all with complementary "sticky" ends. The design strategy for the repetitive HRGP modules combines proven approaches described earlier for the production in E. coli of novel repetitive polypeptide polymers (McGrath et al. [1990] Biotechnol. Prog. 6:188), of a repetitious synthetic analog of the bioadhesive precursor protein of the mussel Mytilus edulis, of a repetitive spider silk protein (Lewis et al. [1996] Protein Express. Purif. 7:400), and of a highly repetitive elastin-like polymer in tobacco [Zhang, X., Urry, D. W., and Daniell, H. "Expression of an environmentally friendly synthetic protein-based polymer gene in transgenic tobacco plants," Plant Cell Reports, 16: 174 (1996)].

[0122] The bas...

example 3

Isolation of Tomato P1 Extensin cDNA Clones

[0138] In order to obtain the tomato P1 extensin signal sequence (i.e., signal peptide), P1 extensin cDNA clones were isolated using oligonucleotides designed after the P1-unique protein sequence: Val-Lys-Pro-Tyr-His-Pro-Thr-Hyp-Val-Tyr-Lys (SEQ ID NO: 51). When present at the N-terminus of a protein sequence, the P1 extensin signal sequence directs the nascent peptide chain to the ER.

example 4

Construction of One Embodiment of an Expression Vector

[0139] pBI121 is an expression vector which permits the high level expression and secretion of inserted genes in plant cells (e.g., tomato, tobacco, members of the genus Solanace, members of the family Leguminoseae, non-graminaceous monocots). pBI121 contains the .sup.35S CaMV promoter, the tobbaco (Nicotiana plumbaginifolia) extensin signal sequence, a EGFP gene, the termination / polyadenylation signal from the nopaline synthetase gene (NOS-ter), a kanamycin-resistance gene (nptII) and the right and left borders of T-DNA to permit transfer into plants by Agrobacterium-mediated transformation.

5TABLE 3 Illustrative HRGP Synthetic Gene Modules 1. MODULES FOR AGP-LIKE SEQUENCES a. The [SP].sub.n Module [SP].sub.n Internal Repeat Oligo's: 5'-TCA CCC TCA CCA TCT CCT TCC CCA TCA CCC (SEQ ID NO:52) GGT AGA GGA AGC GGT AGT GGG AGT GGG AGT-5' (SEQ ID NO:53) The [SP].sub.n 3' & 5' External Linkers for both plasmids are the same as for the G...

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Abstract

A new approach in the field of plant gums is described which presents a new solution to the production of hydroxyproline(Hyp)-rich glycoproteins (HRGPs), repetitive proline-rich proteins (RPRPs) and arabino-galactan proteins (AGPs). The expression of synthetic genes designed from repetitive peptide sequences of such glycoproteins, including the peptide sequences of gum arabic glycoprotein (GAGP), is taught in host cells, including plant host cells.

Description

[0001] The present application is a Continuation-In-Part of U.S. patent application Ser. No. 08 / 897 / 556, filed Jul. 21, 1997.[0002] The present invention relates generally to the field of plant gums and other hydroxyproline-rich glycoproteins, and in particular, to the expression of synthetic genes designed from repetitive peptide sequences.[0003] Gummosis is a common wound response that results in the exudation of a gum sealant at the site of cracks in bark. A. M. Stephen et al., "Exudate Gums", Methods Plant Biochem. (1990). Generally the exudate is a composite of polysaccharides and glycoproteins structurally related to cell wall components such as galactans [G. O. Aspinall, "Plant Gums", The Carbohydrates 2B:522536 (1970)] and hydroxyproline-rich glycoproteins [Anderson and McDougall, "The chemical characterization of the gum exudates from eight Australian Acacia species of the series Phyllodineae." Food Hydrocolloids, 2: 329 (1988)].[0004] Gum arabic is probably the best charac...

Claims

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Application Information

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IPC IPC(8): A01H1/00C07K14/415C07K19/00C12N5/10C12N15/09C12N15/29C12N15/82C12P19/00
CPCC07K14/415C12N15/8242C12N15/8241C07K2319/00
Inventor KIELISZEWSKI, MARCIA J.
Owner OHIO UNIV TECH TRANSFER OFFICE TECH & ENTERPRISE BUILDING
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