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Summer Research Fellowship Programme of India's Science Academies

Cloning of a novel hypothetical gene isolated from moss Physcomitrella patens into pCAMBIA 1302 vector for expression analysis in Nicotiana tabacum

A V Elakkiya

Department of Plant Biotechnology, Centre for Plant Molecular Biology and Biotechnology, Tamil Nadu Agricultural University, Coimbatore 641 003

Dr. Anjan Banerjee

Assoc. Professor & Co-Chair, Biology Div., Indian Institute of Science, Education and Research, Dr. Homi Bhabha Road, Pune 411008.

Abstract

Physcomitrella patens, an extant moss species, has recently emerged as a model organism mainly due to its short life cycle and the ease with which stable transgenic lines can be generated by homologous recombination. The genome of P.patens was recently annotated making it easy to functionally characterize novel genes (Lang et al., 2005). A putative gene SHLF was isolated from the mutant obtained by forward genetics approach in P.patens showing short leaf phenotype. This gene has been hypothesized to be linked to leaf development in moss via an auxin mediated pathway. SHLF contains ER signal in its N-terminal, the function of which remains to be elucidated. In order to observe the resulting phenotype of the translated protein from this gene in Nicotiana, the gene was cloned into binary vector pCAMBIA 1302 under CaMV 35s promoter and over-expression lines in Nicotiana tabaccum was developed. The transgenic over-expression lines can further be used for the functional characterization of this hypothetical gene.

Keywords: mutagenesis, moss, transformation, binary vector, cloning

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