US 2013/0276162 A1 · Gene Constructs Comprising Nucleic Acids That Modulate Chlorophyll Biosynthesis And Uses Thereof

Abstract

The present invention provides isolated nucleic acids encoding Ch1 d synthase, gene constructs comprising the isolated nucleic acids and cells, chloroplasts, plant tissue and whole plants ectopically expressing cyanobacterial Ch1 d synthase. The invention also provides isolated antibodies prepared using recombinant Ch1 d synthase. The antibodies and gene constructs of the invention are used to produce Ch1 d in organisms that do not normally produce Ch1 d, and to modify Ch1 d level in cyanobacterial cells, such as for modifying environmental host range and photosynthetic capacity of organisms in low light and/or red or far-red or near far-red light environments.

Claims39

Dependent claims

A gene construct comprising a Ch1 d synthase gene and one or more origins of replication for maintenance of the gene construct in a cell or chloroplast, wherein said Ch1 d synthase gene comprises a sequence selected from the group consisting of:

(i) the sequence set forth in SEQ ID NO:
11 or a variant thereof comprising a sequence that is degenerate with SEQ ID NO:
11 by virtue of the genetic code and/or that varies from SEQ ID NO:
11 by virtue of a codon usage bias;
(ii) a sequence comprising an open reading frame that encodes the amino acid sequence set forth in SEQ ID NO:
12 or a variant thereof comprising a sequence wherein one or more amino acids of SEQ ID NO:
12 is substituted conservatively for one or more other amino acids in said variant sequence;
(iii) a sequence that is produced by amplification employing one or more primer sequences comprising SEQ ID NO:
14 and SEQ ID NO:
15;
(iv) a sequence having at least about 80% sequence identity to SEQ ID NO:
11;
(v) a sequence comprising an open reading frame that encodes an amino acid sequence having at least about 80% identity to the amino acid sequence set forth in SEQ ID NO:
12;
(vi) a sequence that is produced by amplification employing one or more primer sequences comprising SEQ ID NO:
14 and SEQ ID NO:
15;
(vii) a sequence that is produced by amplification employing primer sequences comprising SEQ ID NO:
18 and SEQ ID NO:
20 or primer sequences comprising SEQ ID NO:
19 and SEQ ID NO:
20;
(viii) a sequence that hybridizes under at least moderate stringency conditions to a sequence that is complementary to (i) or (ii) or fragment thereof comprising at least 10 nucleotides in length;
and
(ix) a sequence that is complementary to a sequence at (i) or (ii) or (iii) or (iv) or (v) or (vi) or (vii) or (viii).

22 more elements

A method of producing Ch1 d in a chloroplast, cell, plant part or plant that normally produces Ch1 a in the absence of Ch1 d, said method comprising expressing a Ch1 d synthase gene in a chloroplast, cell, plant part or plant for a time and under conditions sufficient for Ch1 d to be produced from endogenous Ch1 a in the chloroplast, cell, plant part or plant, wherein said Ch1 d synthase gene comprises a sequence selected from the group consisting of:

(i) the sequence set forth in SEQ ID NO:
11 or a variant thereof comprising a sequence that is degenerate with SEQ ID NO:
11 by virtue of the genetic code and/or that varies from SEQ ID NO:
11 by virtue of a codon usage bias;
(ii) a sequence comprising an open reading frame that encodes the amino acid sequence set forth in SEQ ID NO:
12 or a variant thereof comprising a sequence wherein one or more amino acids of SEQ ID NO:
12 is substituted conservatively for one or more other amino acids in said variant sequence;
(iii) a sequence that is produced by amplification employing one or more primer sequences comprising SEQ ID NO:
14 and SEQ ID NO:
15;
(iv) a sequence having at least about 80% sequence identity to SEQ ID NO:
11;
(v) a sequence comprising an open reading frame that encodes an amino acid sequence having at least about 80% identity to the amino acid sequence set forth in SEQ ID NO:
12;
(vi) a sequence that is produced by amplification employing one or more primer sequences comprising SEQ ID NO:
14 and SEQ ID NO:
15;
(vii) a sequence that is produced by amplification employing primer sequences comprising SEQ ID NO:
18 and SEQ ID NO:
20 or primer sequences comprising SEQ ID NO:
19 and SEQ ID NO:
20;
(viii) a sequence that hybridizes under at least moderate stringency conditions to a sequence that is complementary to (i) or (ii) or fragment thereof comprising at least 10 nucleotides in length;
and
(ix) a sequence that is complementary to a sequence at (i) or (ii) or (iii) or (iv) or (v) or (vi) or (vii) or (viii).

22 more elements

A method of producing Ch1 d in a chloroplast, cell, plant part or plant that does not normally produce a chlorophyll pigment, said method comprising expressing a Ch1 d synthase gene in a chloroplast, cell, plant part or plant in the presence of an amount of exogenous Ch1 a sufficient for Ch1 d to be produced in the chloroplast, cell, plant part or plant, wherein said Ch1 d synthase gene comprises a sequence selected from the group consisting of:

(i) the sequence set forth in SEQ ID NO:
11 or a variant thereof comprising a sequence that is degenerate with SEQ ID NO:
11 by virtue of the genetic code and/or that varies from SEQ ID NO:
11 by virtue of a codon usage bias;
(ii) a sequence comprising an open reading frame that encodes the amino acid sequence set forth in SEQ ID NO:
12 or a variant thereof comprising a sequence wherein one or more amino acids of SEQ ID NO:
12 is substituted conservatively for one or more other amino acids in said variant sequence;
(iii) a sequence that is produced by amplification employing one or more primer sequences comprising SEQ ID NO:
14 and SEQ ID NO:
15;
(iv) a sequence having at least about 80% sequence identity to SEQ ID NO:
11;
(v) a sequence comprising an open reading frame that encodes an amino acid sequence having at least about 80% identity to the amino acid sequence set forth in SEQ ID NO:
12;
(vi) a sequence that is produced by amplification employing one or more primer sequences comprising SEQ ID NO:
14 and SEQ ID NO:
15;
(vii) a sequence that is produced by amplification employing primer sequences comprising SEQ ID NO:
18 and SEQ ID NO:
20 or primer sequences comprising SEQ ID NO:
19 and SEQ ID NO:
20;
(viii) a sequence that hybridizes under at least moderate stringency conditions to a sequence that is complementary to (i) or (ii) or fragment thereof comprising at least 10 nucleotides in length;
and
(ix) a sequence that is complementary to a sequence at (i) or (ii) or (iii) or (iv) or (v) or (vi) or (vii) or (viii).

22 more elements

Dependencies are inferred from the claim text.

Gene Constructs Comprising Nucleic Acids That Modulate Chlorophyll Biosynthesis And Uses Thereof

Life of this patent

Gene Constructs Comprising Nucleic Acids That Modulate Chlorophyll Biosynthesis And Uses Thereof

Life of this patent

Abstract

Claims39