A heterologous expression system for bovine lens transmembrane main intrinsic protein (MIP) in Nicotiana tabacum plants.
We have developed a heterologous expression system for transmembrane lens main intrinsic protein (MIP) in Nicotiana tabacum plant tissue. A native bovine MIP26 amplicon was subcloned into an expression cassette under the control of a constitutive Cauliflower Mosaic Virus promoter, also containing a neomycin phosphotransferase operon. This cassette was transformed into Agrobacterium tumefaciens by triparental mating and used to infect plant tissue grown in culture. Recombinant plants were selected by their ability to grow and root on kanamycin-containing media. The presence of MIP in the plant tissues was confirmed by PCR, RT-PCR and immunohistochemistry. A number of benefits of this system for the study of MIP will be discussed, and also its application as a tool for the study of heterologously expressed proteins in general.
de Peyer
O S
OS
Division of Cell and Molecular Biology, School of Animal and Microbial Sciences, The University of Reading, Whiteknights, Reading, Berkshire, UK. astrobiologist@hotmail.com
Wetten
A C
AC
Dunwell
J M
JM
Crabbe
M J
MJ
D000818:Animals; D020346:Aquaporins; D002417:Cattle; D003001:Cloning, Molecular; D005136:Eye Proteins / Q000235:genetics*; D015870:Gene Expression; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins; D030821:Plants, Genetically Modified / Q000235:genetics*; D010947:Plants, Toxic; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014026:Nicotiana / Q000235:genetics*
D020346:Aquaporins; D005136:Eye Proteins; D008562:Membrane Glycoproteins; C030328:aquaporin 0