Membrane Transporter Expression Facility
What is the membrane transporter expression facility (MTEF)?
The MTEF enables expression of genes (plant or animal) in a cell system (e.g. Xenopus oocytes, plant cell line or protoplast, animal cell line) that can be robotically screened for various membrane transport activities and has applications in drug discovery and protein engineering. The facility includes both manual and automated electrophysiology equipment for voltage clamp or flux experiments (e.g. patch clamp or TEVC systems, the robocyte and MIFE).
The core of the facility is the ‘robocyte’ that was obtained in a recent Australian Research Council (LIEF) grant. The robocyte is a high throughput Xenopus oocyte mRNA injecting and voltage clamp apparatus that is designed to screen 96 Xenopus oocytes for transport protein activity in a consecutive manner. A solution handling robot is combined to allow screening of pharmaceuticals and ion/solute responses.
Imaging and TEVC system for initial screening
The MTEF incorporates a Xenopus frog colony maintained by the University of Adelaide Animal Housing facilities, plus expertise in surgically removing oocytes from frogs without sacrificing the animals. The MTEF has a dedicated research associate engaged to harvest oocytes and setting up the robocyte for researchers expressing a very broad array of plant and animal genes encoding membrane transporters and associated proteins.
The MTEF will enable the functional characterisation of newly discovered transporter genes and enable researchers to screen pharmaceuticals for activity on the transporter protein. The facility will also provide advice on how to prepare genes for expression and determine the transport function of expressed proteins. It is possible to characterise non-electrogenic and electrogenic transporters, as well as aquaporin genes.
A number of patents have been born from work at the MTEF, plus a great deal of publications over recent years.
Please contact us if you are interested in using these facilities.
Recent Selected publications
Munns RJ, James RA, Xu B, Athman A, Conn SJ, Jordans C, Byrt CS, Hare RA, Tyerman SD, Tester M, Plett D and Gilliham M (2012) Grain yield of modern wheat on saline soils is improved by ancestral HKT gene. Nature Biotechnology 30:360-364
Wang C, Huang W, Ying Y, Li S, Secco D, Tyerman S, Whelan J and Shou H (2012) Functional characterization of the rice SPX-MFS family reveals a key role of OsSPX-MFS1 in controlling phosphate homeostasis in leaves. New Phytologist doi: 10.1111/j.1469-8137.2012.04227.x
Kotur Z, Mackenzie N, Ramesh S, Tyerman SD, Kaiser BN and Glass ADM (2012) Nitrate transport capacity of the Arabidopsis thaliana NRT2 family members and their interactions with AtNAR21. New Phytologist. (Accepted 31/1/2012)
Secco D, Wang C, Arpat BA, Wang Z, Poirier Y, Tyerman SD, Wu P, Shou H and Whelan J (2012) The emerging importance of the SPX domain-containing proteins in phosphate homeostasis. New Phytologist. DOI: 101111/j1469-8137201104002x
Michard E, Lima PT, Borges F Silva AC, Carvalho JE, Gilliham M, Liu L-H, Obermeyer G, Feijó JA (2011) Glutamate-Receptor-like genes control pollen tube caclium ion influx and morphogenesis. Science 6028: 434-437. Cover image
Conn SJ, Gilliham M, Athman A, Schreiber AS, Baumann U, Moller I, Cheng N-H, Stancombe MA, Hirschi KD, Webb AAR, Burton R, Kaiser BN, Tyerman SD and Leigh RA (2011) Cell-specific vacuolar calcium storage mediated by AtCAX1 regulates apoplastic calcium concentration, gas exchange and plant productivity. Plant Cell 23:240-257
Preuss CP, Huang CY and Tyerman SD (2011) Proton-coupled high-affinity phosphate transport revealed from heterologous characterization in Xenopus of barley-root plasma membrane transporter, HvPHT1;1. Plant Cell and Environment. 34: 681-689
Gruber BD, Ryan PR, Richardson AE, Tyerman SD, Ramesh S, Hebb DH, Howitt SM and Delhaize E (2010) HvALMT1 from barley is involved in the transport of organic anions. Journal of Experimental Botany. 61:1455-67
Preuss CP, Huang, CY Gilliham, M and Tyerman SD 2010 Channel-like characteristics of the low-afï¬ï‚Ânity barley phosphate transporter PHT1;6 when expressed in Xenopus oocytes. Plant Physiology. 152: 1431-41
Schnurbusch T, Hayes J, Hrmova M, Baumann U, Ramesh SA, Tyerman SD, Langridge P and Sutton T (2010) Boron toxicity tolerance in barley through reduced expression of the multifunctional aquaporin HvNIP2;1. Plant Physiology. 153:1706-15
Møller IS, Gilliham M, Jha D, Mayo GM, Roy SJ, Coates JC, Haseloff J, Tester M (2009) Shoot sodium ion exclusion and increased salinity tolerance engineered by cell type-specific manipulation of sodium ion transport in Arabidopsis. Plant Cell 21: 2163-2178.
Roy SJ, Gilliham M, Berger B, Essah PA, Cheffings C, Miller AJ, Widdowson L, Davenport RJ, Liu L-H, Skynner MJ, Davies JM, Richardson P, Leigh RA, Tester M (2008) Investigating glutamate receptor-like gene co-expression in Arabidopsis thaliana. Plant, Cell and Environment 31: 861-871.
Shelden MC, Howitt SM, Kaiser BN and Tyerman SD (2009) Identification and functional characterisation of aquaporins in the grapevine, Vitis Vinifera. Functional Plant Biology. 36:1065-78 (Awarded ECR Best Paper 2010)
Vandeleur RK, Mayo G, Shelden MC, Gilliham M, Kaiser BN and Tyerman SD (2009) The role of plasma membrane intrinsic protein aquaporins in water transport through roots: diurnal and drought stress responses reveal different strategies between isohydric and anisohydric cultivars of grapevine. Plant Physiology. 149:445-60
Fitzpatrick KL, Tyerman SD and Kaiser BN (2008) Molybdate transport through the plant sulfate transporter SHST1. FEBS Letters. 582:1508-13
Zhang WH, Ryan PR, Sasaki T, Yamamoto Y, Sullivan W and Tyerman SD (2008) Characterization of the TaALMT1 protein as an Aluminum-activated anion channel in transformed tobacco (Nicotiana tabacum L) cells. Plant and Cell Physiology. 49:1316-30
Zhou Y, Setz N, Niemietz C, Qu H, Offler CE, Tyerman SD and Patrick JW (2007) Aquaporins and unloading of phloem-imported water in coats of developing bean seeds. Plant Cell and Environment. 30:1566-77