| CPC A01K 67/0339 (2013.01) [C12N 15/8509 (2013.01); C12Q 1/6888 (2013.01); A01K 2217/072 (2013.01); A01K 2217/15 (2013.01); A01K 2217/203 (2013.01); A01K 2217/206 (2013.01); A01K 2217/30 (2013.01); A01K 2227/706 (2013.01); A01K 2267/02 (2013.01); C12N 2800/90 (2013.01); C12N 2830/006 (2013.01); C12N 2830/008 (2013.01); C12N 2830/15 (2013.01); C12N 2840/007 (2013.01); C12N 2840/44 (2013.01); C12N 2840/75 (2013.01); C12N 2999/007 (2013.01); C12Q 2600/158 (2013.01)] | 4 Claims |
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1. An isolated polynucleotide construct, wherein said polynucleotide construct comprises a first and a second gene expression system forming a first and second positive feedback loop to be expressed in an insect:
i) the first gene expression system comprises the following components: a first dominant lethal gene operably linked to a first promoter, a gene encoding a first activating transcription factor, and a first splice control sequence,
ii) the second gene expression system comprises the following components: a second dominant lethal gene operably linked to a second promoter, a gene encoding a second activating transcription factor, and a second splice control sequence, wherein:
each of said activating transcription factors is capable of activating at least one of said promoters, and both of said promoters are activated when both of said transcription factors are expressed,
each of the first and second splice control sequences mediates female-specific expression of the first and second dominant lethal genes in female insects, respectively, by alternative splicing, and such expression leads to the death of said female insects at an early stage of development,
the transactivation activity of each of the first and second activating transcription factors is repressible by a first and a second exogenous control factor, respectively, wherein said first exogenous control factor is the same as or different from said second exogenous control factor, and
each of said components of said first gene expression system are the same as or different from said components of said second gene expression system;
wherein one of the first and second gene expression systems comprises a VP16 encoding sequence and a third promoter operably linked to the VP16 sequence, wherein the activating transcription factor capable of activating the first or second promoter of said gene expression system is also capable of activating the third promoter, wherein:
the first dominant lethal gene is tTAV (SEQ ID NO: 26), the first activating transcription factor is the tTAV gene product, the first promoter is Hsp70, the first splice control sequence is Cctra,
the second dominant lethal gene is tTAV3 (SEQ ID NO: 28), the second activating transcription factor is the tTAV3 gene product, the second promoter is srya, and the second splice control sequence is Bztra,
the first gene expression system further comprises a first enhancer associated with the first promoter, wherein the first enhancer is tetOx7,
the second gene expression system further comprises a second enhancer associated with the second promoter, wherein the second enhancer is tetOx14, the polynucleotide construct further comprises a third dominant lethal gene operably linked to a third promoter, the third promoter being associated with the second enhancer, wherein the third dominant lethal gene is VP16 and the third promoter is Hsp70, wherein the second promoter is associated with one end of the second enhancer and the third promoter is associated with the other end of the second enhancer, and
the polynucleotide construct further comprises a first genetic marker, which is DsRed2.
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