Transactivation of a target gene using a suppressor tRNA in transgenic tobacco plants

The abundance of tRNAs, together with their central role in translation, has generated considerable interest in the use of tRNA genes for biotechnological applications. One such application is the use of suppressor tRNAs to transactivate target genes containing premature stop codons. Previous work has shown that such systems can work in transient expression experiments in plant protoplasts; here these experiments are extended to show that suppression of stop codons can occur in whole plants. Transgenic tobacco plants homozygous for a modified tRNA^Leu gene expressing a strong amber suppressor tRNA, and plants carrying a β-glucuronidase (gus) gene inactivated by a premature amber stop codon have been obtained. When the two types of plants are crossed, many of the F₁ hybrids show significant GUS activity. The GUS activity is dependent on the presence of both the suppressor tRNA gene and the gus gene. Tobacco plants carrying the suppressor tRNA gene are phenotypically normal, fertile and the gene shows normal Mendelian inheritance. The potential applications of such a system are discussed.     

Influence of kinematic analysis methods on detecting ankle and subtalar joint instability

Patients with subtalar joint instability may be misdiagnosed with ankle instability, which may lead to chronic instability at the subtalar joint. Therefore, it is important to understand the difference in kinematics after ligament sectioning and differentiate the changes in kinematics between ankle and subtalar instability. Three methods may be used to determine the joint kinematics; the Euler angles, the Joint Coordinate System (JCS) and the helical axis (HA). The purpose of this study was to investigate the influence of using either method to detect subtalar and ankle joints instability. 3D kinematics at the ankle and subtalar joint were analyzed on 8 cadaveric specimens while the foot was intact and after sequentially sectioning the anterior talofibular ligament (ATFL), the calcaneofibular ligament (CFL), the cervical ligament and the interosseous talocalcaneal ligament (ITCL). Comparison in kinematics calculated from sensor and anatomical landmarks was conducted as well as the influence of Euler angles and JCS rotation sequence (between ISB recommendation and previous research) on the subtalar joint. All data showed a significant increase in inversion when the ITCL was sectioned. There were differences in the data calculated using sensors coordinate systems vs. anatomic coordinate systems. Anatomic coordinate systems were recommended for these calculations. The Euler angle and JCS gave similar results. Differences in Euler angles and JCS sequence lead to the same conclusion in detecting instability at the ankle and subtalar joint. As expected, the HA detected instability in plantarflexion at the ankle joint and in inversion at the subtalar joint.