Insulin-like gene of prawns and uses thereof
AssigneeBen-Gurion University of the Negev Research and Development Authority
InventorsAmir Sagi, Tomer Ventura
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Abstract
Nucleic acid molecules encoding an insulin-like factor of the androgenic gland of the freshwater prawn Macrobrachium rosenbergii (M. rosenbergii) are disclosed. Also disclosed are methods of silencing the expression of the insulin-like factor gene in decapod crustaceans order, particularly in M. rosenbergii, useful for producing male monosex populations.
Claims5
A double stranded RNA (dsRNA) molecule, wherein the sequence of one strand of said dsRNA molecule is at least 80% complementary to a fragment of SEQ ID NO:1, wherein said one strand of said dsRNA is 100 to 700 nucleotides in length.
A double stranded RNA (dsRNA) molecule, wherein said dsRNA molecule is obtained by a process comprising the steps of:
- (a) obtaining an antisense PCR product of a template comprising SEQ ID NO:
- 2 and the primers of SEQ ID NOs:
- 23 and 24;
- (b) obtaining a sense PCR product of a template comprising SEQ ID NO:
- 2 and the primers of SEQ ID NOs:
- 21 and 22;
- (c) transcribing said antisense PCR product and said sense PCR product of step (a) and step (b),
- (d) hybridizing the antisense RNA and the sense RNA obtained in step (c);
- thereby, obtaining said dsRNA molecule.
A DNA construct for generating a double stranded RNA capable of down regulating the expression of an insulin-like factor gene of M. rosenbergii, wherein the DNA construct comprises a promoter operably linked to a nucleic acid molecule encoding an RNA sequence that forms a double stranded RNA, the nucleic acid molecule comprises:
- (i) a first nucleotide sequence having at least 80% sequence identity to SEQ ID NO:1 or a fragment thereof;
- and
- (ii) a second nucleotide sequence having at least 80% sequence identity to a complementary sequence of SEQ ID NO:1 or a fragment thereof,
- wherein the RNA transcribed from the DNA construct down regulates the expression of the insulin-like factor gene, wherein said fragment thereof comprises 100 to 700 nucleotides.
Dependencies are inferred from the claim text.