Large Scale Biology Corporation

Novici Biotech LLC

Other

Wayne P Fitzmaurice

John A Lindbo

Hal S Padgett

Gregory P Pogue

Laurence K Grill

Monto H Kumagai

Andrew A Vaewhongs

Gary Mark Hellmann

Mark L Smith

Guy R Della-Cioppa

Guy Richard della-Cioppa

Fakhrieh S Vojdani

We describe here an in vitro method of increasing complementarity in a heteroduplex polynucleotide sequence. The method uses annealing of opposite strands to form a polynucleotide duplex with mismatches. The heteroduplex polynucleotide is combined with an effective amount of enzymes having strand cleavage activity, 3' to 5&#x2032; exonuclease activity, and polymerase activity, and allowing sufficient time for the percentage of complementarity to be increased within the heteroduplex. Not all heteroduplex polynucleotides will necessarily have all mismatches resolved to complementarity. The resulting polynucleotide is optionally ligated. Several variant polynucleotides result. At sites where either of the opposite strands has templated recoding in the other strand, the resulting percent complementarity of the heteroduplex polynucleotide sequence is increased. The parent polynucleotides need not be cleaved into fragments prior to annealing heterologous strands. Therefore, no reassembly is required.

Method of increasing complementarity in a heteroduplex

We describe here an in vitro method of redistributing sequence variations between non-identical polynucleotide sequences, by making a heteroduplex polynucleotide from two non-identical polynucleotides; introducing a nick in one strand at or near a base pair mismatch site; removing mismatched base(s) from the mismatch site where the nick occurred; and using the opposite strand as template to replace the removed base(s) with bases that complement base(s) in the first strand. By this method, information is transferred from one strand to the other at sites of mismatch.

Population of polynucleotide sequence variants

The present invention provides nucleic acid sequences having an altered viral movement protein and 126/183 kDa replicase proteins further characterized in its ability tostabilize a transgene contained in a virus that expresses the altered movement protein. The present invention also provides viral vectors expressing the altered movement protein, cells transformed with the vectors, and host plants infected by the viral vectors.

Viral expression vectors

We describe here restriction endonucleases and their uses. Restriction endonucleases are useful in finding single nucleotide polymorphisms. They are also useful in an in vitro method of redistributing sequence variations between non-identical polynucleotide sequences.

Mismatch endonucleases and methods of use

The present invention provides nucleic acid sequences having an altered viral movement protein and 126/183 kDa replicase proteins further characterized in its ability to stabilize a transgene contained in a virus that expresses the altered movement protein. The present invention also provides viral vectors expressing the altered movement protein, cells transformed with the vectors, and host plants infected by the viral vectors.

The present invention relates to a method for using viral vectors to bear populations of sequence variants and using plant hosts to select the sequences that exhibit the desired traits.

Method for expressing a library of nucleic acid sequence variants and selecting desired traits

The present invention relates to novel interspecific hybrid seeds and plants and to a method of producing interspecific Nicotiana hybrids having enhanced properties for biomass and the production of recombinant proteins using a viral vector system.

Interspecific nicotiana hybrids and their progeny

DNA sequences isolated from Nicotiana species (e.g., Nicotiana tabacum and Nicotiana benthamiana) and encoding polypeptides having enzymatic activity for producing phytoene, and the polypeptides encoded, are provided.

DNA sequences encoding enzymes useful in phytoene biosynthesis

DNA sequences isolated from Nicotiana species (e.g., Nicotiana benthamiana and Nicotiana tabacum) have SEQ ID NOS: 1, 3, 5 and 7. The DNA sequences encode polypeptides having enzymatic activity for producing zeta-carotene. The polypeptides are referred to as phytoene desaturase.

Growing community of inventors

Vacaville, CA, United States of America

Wayne P Fitzmaurice