Pal Maliga Professor Rutgers University Department of Genetics Waksman Institute Piscataway. NJ 08854-0759 (732) 445-5329 FAX - 5735 maliga@waksman.rutgers.edu

Molecular genetics of plastids. plastid engineering. regulation of plastid gene expression

The plastids of higher plants are genetically semi-autonomous: they have their own genome. transcription and translation machinery. Our long-term interest is to understand how coordinated expression of plastid and nuclear genes contribute to plastid function and plant development. We extensively use transgenic technology. which was pioneered for higher plant plastids in this laboratory. Our model plants are tobacco. Arabidopsis and tomato. (1) We study the multisubunit (PEP) and phage-type (NEP) plastid RNA polymerases: promoter architecture. recognition. polymerase composition and regulation of plastid gene expression utilizing a combination of in vitro biochemical and in vivo transgenic approaches. (2) We are working on the implementation of transplastomic technology in Arabidopsis thaliana. This research utilizes plant cell culture and phage recombinases that are suitable for the insertion of transforming DNA. marker gene excision. and to probe plastid gene function. (3) A new area is the study of the role of plastid proteases in plant development. (4) We are interested in the biotechnological applications of transplastomic technology. including expression of recombinant proteins for biomedical and industrial uses.

Selected Publications

Lenzi P, Scotti N, Alagna F, Tornesello ML, Pompa A, Vitale A, De Stradis A, Monti L, Grillo S, Buonaguro FM, Maliga P, Cardi T. (2008) Translational fusion of chloroplast-expressed human papillomavirus type 16 L1 capsid protein enhances antigen accumulation in transplastomic tobacco. Transgenic Res. May 20. [Epub ahead of print]

Sharwood RE, von Caemmerer S, Maliga P, Whitney SM. (2008) The catalytic properties of hybrid Rubisco comprising tobacco small and sunflower large subunits mirror the kinetically equivalent source Rubiscos and can support tobacco growth. Plant Physiol. 146(1):83-96.

Lutz KA, Azhagiri AK, Tungsuchat-Huang T, Maliga P. (2007) A guide to choosing vectors for transformation of the plastid genome of higher plants. Plant Physiol. 145(4):1201-10.

Azhagiri AK, Maliga P. (2007) Exceptional paternal inheritance of plastids in Arabidopsis suggests that low-frequency leakage of plastids via pollen may be universal in plants. Plant J. 52(5):817-23.

Lutz KA, Maliga P. (2007) Transformation of the plastid genome to study RNA editing. Methods Enzymol. 424:501-18.

Svab Z, Maliga P. (2007) Exceptional transmission of plastids and mitochondria from the transplastomic pollen parent and its impact on transgene containment. Proc Natl Acad Sci U S A. 104(17):7003-8.

Lutz, K.A., Svab, Z. and Maliga, P. (2006) Construction of marker-free transplastomic tobacco using the CRE-loxP site-specific recombination system. Nature Protocols. 1(2): 900-910.

Tungsuchat, T., Kuroda, H., Narangajavana, J. and Maliga, P. (2006) Gene activation in plastids by the CRE site-specific recombinase. Plant Mol. Biol. 61(4-5): 711-718.

Chakrabarti, S.K., Lutz, K.A., Lertwirijawong, B., Svab, Z. and Maliga, P. (2006) Expression of the cry9Aa2 Bt gene in tobacco chloroplasts confers extreme resistance to potato tuber moth. Transgenic Research 15: 485-492.

Lutz KA. Bosacchi MH. Maliga P. (2006) Plastid marker-gene excision by transiently expressed CRE recombinase. Plant J. 45(3):447-56.

Maliga P. (2005) New vectors and marker excision systems mark progress in engineering the plastid genome of higher plants. Photochem Photobiol Sci. 4(12):971-6.

Tregoning JS. Clare S. Bowe F. Edwards L. Fairweather N. Qazi O. Nixon PJ. Maliga P. Dougan G. Hussell T. (2005) Protection against tetanus toxin using a plant-based vaccine. Eur J Immunol. 35(4):1320-6.

Maliga. P. (2004) Plastid transformation in higher plants. Annu. Rev. Plant Biol. 55:289-313.

Corneille. S.. Lutz. K.A.. Azhagiri. A.K. and Maliga. P. (2003) Identification of functional lox sites in the plastid genome. Plant J. 35: 753-762.

Kuroda. H. and Maliga. P. (2003) The plastid clpP1 gene is essential for plant development. Nature 425: 86-89.

Maliga. P. (2003) Mobile plastid genes. Nature 422: 31-32.

Maliga. P. (2003) Progress towards commercialization of plastid transformation technology. Trends in Biotechnology. 21:20-28.

Suzuki. J.. Sriraman. P. Svab. Z. and Maliga. P. (2003) Unique architecture of the plastid rRNA operon recognized by the multisubunit RNA polymerase in tobacco and other higher plants. Plant Cell 15: 195-215.

Tregoning. J.. Nixon. Kuroda. H.. Svab. Z.. Clare. S.. Bowe. F.. Fairweather. N.. Y. Ytterberg. K.J. vanWijk. Dougan. G. and Maliga. P. (2003) Expression of tetanus toxin fragment C in tobacco chloroplasts. Nucleic Acids Res. 31: 1174-1179.

Maliga. Pal (2002) Engineering the plastid genome of higher plants. Curr. Opin. Plant Biol. 5: 164-172.

Lutz. K. and Maliga. P. (2001) Lack of conservation of editing sites in mRNA that encode subunits of the ND(P)H dehydrogenase complex in plastids and mitochondria of Arabidopsis thaliana. Curr. Genet. 40:214-219.

Maliga. P. (2001) Plastid engineering bears fruit. Nat. Biotechnol. 19: 826-827

Corneille. S.. Lutz. K.. Svab. Z. and Maliga. P. (2001) Efficient elimination of selectable marker genes from the plastid gnome by the CRE-lox site-specific recombination system. Plant J. 27: 171-178.

Lutz. K.A.. Knapp. J.E. and Maliga. P. (2001) Expression of bar in the plastid genome confers herbicide resistance. Plant Physiol. 125: 1585-1590.

Kuroda. H. and Maliga. P. (2001) Complementarity of the 16S rRNA penultimate stem with sequences downstream of the AUG destabilizes the plastid mRNAs. Nucleic Acids Res. 29: 970-975.

Kuroda. H. and Maliga. P. (2001) Sequences downstream of the translation initiation codon are important determinants of translation efficiency in chloroplasts. Plant Physiol. 125: 430-436.