Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Utility Patent
1997-09-19
2001-01-02
Achutamurthy, Ponnathapu (Department: 1652)
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
C435S006120, C530S350000, C530S370000
Utility Patent
active
06169174
ABSTRACT:
TECHNICAL FIELD
The present invention relates to a novel gene of cotton plant. In particular, the present invention relates to a gene which is transcribed in a large amount in fiber cells during cotton fiber formation. In particular, the present invention relates to a gene coding for an amino acid sequence having zinc finger motif, the zinc finger motif being one of characteristic sequences of transcription factors.
BACKGROUND ART
Cotton plant occupies an extremely important position as a plant for raw material supply in the textile industry as a matter of course, as well as in the paper-manufacturing industry and the food industry. Therefore, improvement of cotton plant is extensively performed. Efforts to improve cotton plant are made in various viewpoints, such as those for addition and improvement of various characteristics, including, for example, those concerning increase in oil content and improvement in disease resistance, cold resistance, drought resistance, salt tolerance, herbicide resistance, high-yielding ability, early harvest characteristics, and fiber characteristics.
Especially, improving procedures, which have been impossible by using the conventional method, are being carried out by regulating gene expression in a plant by using genetic engineering techniques. When expression of a foreign or endogeneous gene is regulated in the plant in accordance with such a method, it is desirable that the gene to be introduced has been previously isolated, or information has been previously obtained for a sequence of the gene for which expression is regulated, a sequence of an upstream region therefrom, a tissue in which the gene is expressed, and a timing or stage of expression.
As for cotton plant, a small number of genes and little genetic information are still known to be used for the techniques as described above. In such circumstances, although several study reports have been made for what kinds of genes are transcribed, translated, and expressed to what extent during fiber formation in cotton plant (M. E. John et al.,
Proc. Natl. Acad. Sci. USA,
89, 5769 (1992); T. A. Wilkins,
Plant Physiology,
102, 679 (1993); and Din-Pow Ma et al.,
BBA,
1257, 81 (1995)), the knowledge is still insufficient. Especially, there has not been isolated any transcription factor-like gene which should be expressed in a large amount in fiber cells during cotton fiber formation.
SUMMARY OF THE INVENTION
An object of the present invention is to provide a gene which regulates the fiber formation mechanism in cotton plant, and which is used for industrially useful cotton plant through various improving procedures. Another object of the present invention is to provide a gene possibly coding for a transcription factor, expressed in a large amount in fiber cells of cotton plant during cotton fiber formation.
It is known that the transcription factor binds to a specific DNA sequence on genome, and thus it regulates transcription of a specified gene. Owing to this mechanism, the timing or stage and the location of expression of respective genes are controlled in an organized manner.
Provided that a transcription factor is expressed in a large amount in cotton plant fiber cells during cotton fiber formation, it is considered that expression of a gene of such a transcription factor is regulated in fiber cells during fiber formation, and consequently the organ of cotton fiber is formed by controlling the transcription of a gene which specifically interacts with the transcription factor, for its timing and location, i.e., during fiber formation in fiber cells.
Therefore, it is possible to control the expression of the gene which specifically interacts with the transcription factor, by controlling the expression itself of the gene coding for the transcription factor. Thus it is possible to modify the character of fiber, create fiber not known until the present, and create a seed having no fiber. In that way, it is considered that cotton plant useful for the fiber industry, the paper-manufacturing industry, and the food industry can be provided.
A gene, which is expressed in a large amount in cotton plant fiber cells during cotton fiber formation and has a transcription factor-like motif, may not actually have the function of a transcription factor. It is considered that expression of even such a gene affects the fiber formation mechanism. Accordingly, it can be expected that any effect is exerted during cotton fiber formation by controlling expression of the gene, and any improved effect appears on fiber.
Further, it is sufficiently conceivable that genes coding for polyamino acids having the zinc finger motif exist in the cotton plant gene, other than the gene of the present invention. Therefore, it is possible to isolate various genes coding for polyamino acids having the zinc finger motif, including the gene of the present invention, by using, as a probe, a sequence portion coding for the zinc finger motif. Thus it is expected that transcription of many genes can be controlled in cotton plant.
Further, a transcriptional regulatory region existing in an upstream region of the genome may be isolated by using, as a probe, a part or all of a sequence of a coding region of such a gene. An arbitrary gene may be ligated at an downstream position from the transcription regulatory region, and the obtained DNA may be introduced into the cotton gene. Thus it is also possible to express the gene in a large amount in fiber cells during fiber formation. Such an approach may be industrially useful.
The present inventors prepared a cDNA library from cotton plant. The present inventors randomly selected a large amount of clones therefrom to subsequently determine their nucleotide sequences so that homology search was performed between the determined nucleotide sequences and known gene sequences included in a gene data base. In general, a method is used for cloning a gene, in which a part or all of a gene sequence having been isolated from another organism is used as a probe to perform screening from a library. However, in the present invention, homology search was performed between the nucleotide sequences of the clones randomly selected from the cDNA library and the known gene sequences in the data base. Thus it was found that a cotton plant gene was present, having high homology to a sunflower gene SF3 considered to correspond to a transcription factor specifically expressed in sunflower pollen and having two zinc finger motifs (Rachel Baltz et al.,
The Plant Journal,
2(5), 713-721 (1992)). Further, it was found that the gene was expressed in a large amount in fiber cells, by means of northern analysis for the cotton plant gene. Thus the present invention has been completed.
Namely, the present invention lies in a DNA coding for a protein as defined in the following (A) or (B):
(A) a protein having an amino acid sequence shown in SEQ ID NO: 2; or
(B) a protein comprising an amino acid sequence in which one or several amino acids are deleted, substituted, inserted, or added with respect to the sequence shown in SEQ ID NO: 2, and comprising a polyamino acid having a zinc finger motif.
The nucleotide sequence of the DNA of the present invention is specifically exemplified by DNA as defined in the following (a) or (b):
(a) a DNA having a sequence of nucleotide numbers of 134 to 757 in SEQ ID NO: 1; or
(b) a DNA which is hybridizable with the DNA as defined in the foregoing item (a) under a stringent condition and which codes for a protein comprising a polyamino acid having a zinc finger motif.
The DNA is a gene which is expressed in a large amount in fiber cells during cotton fiber formation. In particular, the DNA is a gene which codes for an amino acid sequence having a zinc finger motif as one of characteristic sequences of the transcription factor. The gene is herein referred to as “the gene of the present invention”, if necessary.
In another aspect of the present invention, there is provided a DNA probe as a probe for hybridization, used to isolate a transcription regulatory region of a gene expressed du
Aotsuka Satoshi
Hasegawa Osamu
Takenishi Soichiro
Uchimiya Hirofumi
Achutamurthy Ponnathapu
Jones & Askew LLP
Mayhew Bradley S.
Nisshinbo Industries Inc.
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