Characterization and plant expression of glyphosate-tolerant enolpyruvylshikimate phosphate synthase

Abstract BACKGROUND: Glyphosate tolerance is a dominant trait in modern biotech crops. RESULTS: A gene encoding a glyphosate-tolerant EPSP synthase (aroA 1398 ) from bacterial strain ATX1398 was cloned and characterized. The protein is initiated at a GTG translational start codon to produce a protein that provides robust glyphosate resistance in Escherichia coli (Mig) Cast & Chalm. The aroA 1398 protein was expressed and purified from E. coli, and key kinetic values were determined (K i = 161 µM; K m (PEP) = 11.3 µM; k cat = 28.3 s −1 ). The full-length enzyme is 800-fold more resistant to glyphosate than the maize EPSP synthase while retaining high affinity for the substrate phosphoenol pyruvate. To evaluate further the potential of aroA 1398 , transgenic maize events expressing the aroA 1398 protein were generated. T 0 plants were screened for tolerance to glyphosate sprays at 1.3× commercial spray rates, and T 1 plants were selected that completely resisted glyphosate sprays at 1×, 2× and 4× recommended spray rates in field trials. CONCLUSION: These data suggest that aroA 1398 is a suitable candidate for conferring glyphosate tolerance in transgenic crop plants

Behavioural and functional characterization of Kv10.1 (Eag1) knockout mice

Kv10.1 (Eag1), member of the Kv10 family of voltage-gated potassium channels, is preferentially expressed in adult brain. The aim of the present study was to unravel the functional role of Kv10.1 in the brain by generating knockout mice, where the voltage sensor and pore region of Kv10.1 were removed to render non-functional proteins through deletion of exon 7 of the KCNH1 gene using the ‘3 Lox P strategy’. Kv10.1-deficient mice show no obvious alterations during embryogenesis and develop normally to adulthood; cortex, hippocampus and cerebellum appear anatomically normal. Other tests, including general health screen, sensorimotor functioning and gating, anxiety, social behaviour, learning and memory did not show any functional aberrations in Kv10.1 null mice. Kv10.1 null mice display mild hyperactivity and longer-lasting haloperidol-induced catalepsy, but there was no difference between genotypes in amphetamine sensitization and withdrawal, reactivity to apomorphine and haloperidol in the prepulse inhibition tests or to antidepressants in the haloperidol-induced catalepsy. Furthermore, electrical properties of Kv10.1 in cerebellar Purkinje cells did not show any difference between genotypes. Bearing in mind that Kv10.1 is overexpressed in over 70% of all human tumours and that its inhibition leads to a reduced tumour cell proliferation, the fact that deletion of Kv10.1 does not show a marked phenotype is a prerequisite for utilizing Kv10.1 blocking and/or reduction techniques, such as siRNA, to treat cancer

Contribución al desarrollo de sistemas de transformación genética para la obtención de plantas resistentes a la broca del café hypothenemus hampei (Ferrari)

IP 2251-12-620-96v.1. Contribucion al desarrollo de sistemas de transformaciongenetica para la obtencion de plantas resistentes a la broca del cafe hypothenemus hampei(ferrari) /Jose Ricardo Acula Zornosa -- v.2. Aislamiento y cultivo de protoplastos a partir de suspensiones celulares dedos genotipos de : coffea arabica var Colombia / Harold Mondragon Alvarez.ARTICULO(S) EN REVISTA: Busqueda de secuencias homologas agenesde resistencia a insectos en el genoma de;(2003); p. -- ISSN 00221198 -- Coffee seeds contain 11s storageproteins /Ricardo Acuña ... [et al.] -- En;Physiologia Plantarium. -- Vol. 105. (1999); p. 122-131. -'- isSN319317'-- Optimised dna delivery into;Coffea arabica suspension culture cells by particle bombardment./ AdrianaRosillo ... [et al.] -- En: Plant;Cell, Tissue and Organ Culture. -- Vol. 74 (2003); p. 45-49 --ISSN 0167-6857.;Coffea arabica L., Cv Colombia / Elsa Leonor Alvarez-Mendez ...[et al.] -- En Cenicafe. -- vol. 53, no.