Performance of New Muchamiel Tomato Lines with Virus Resistance Genes Grafted onto Two Commercial Rootstocks

Tomato landraces are regaining interest in Spain because their great fruit quality and value in popular gastronomy. Muchamiel is a traditional tomato variety grown in SE Spain that has been recently improved by the CIAGRO-UMH Tomato Breeding Group, resulting in several lines and hybrids with genetic resistances to virus and most of the original Muchamiel genome. In the current study, two hybrids and one pure line from CIAGRO-UMH and a commercial Muchamiel were grown under conventional conditions to evaluate three different grafting treatments: non-grafting and grafting onto the commercial Beaufort and Maxifort rootstocks. The yield parameters and fruit quality were assessed, and a sensory analysis was performed to evaluate the behavior of every scion/rootstock combination. Overall, significantly worse yield and fruit number in Maxifort-grafted plants were reported; as well as a slight reductions in SSC, fructose, and sucrose; and significant effects on few sensory traits. Instead, Beaufort-grafted plants showed no reduced yield, whereas no differences were reported between grafting treatments in fruit weight, TA, and acid profile, as well as in most of flavor and texture sensory parameters. These results suggest that Muchamiel/Beaufort combination could be suitable under unfavorable conditions, while Maxifort do not seem to provide agronomic nor quality benefits.

Development of a universal nanobody-binding Fab module for fiducial-assisted cryo-EM studies of membrane proteins

With conformation-specific nanobodies being used for a wide range of structural, biochemical, and cell biological applications, there is a demand for antigen-binding fragments (Fabs) that specifically and tightly bind these nanobodies without disturbing the nanobody–target protein interaction. Here, we describe the development of a synthetic Fab (termed NabFab) that binds the scaffold of an alpaca-derived nanobody with picomolar affinity. We demonstrate that upon complementary-determining region grafting onto this parent nanobody scaffold, nanobodies recognizing diverse target proteins and derived from llama or camel can cross-react with NabFab without loss of affinity. Using NabFab as a fiducial and size enhancer (50 kDa), we determined the high-resolution cryogenic electron microscopy (cryo-EM) structures of nanobody-bound VcNorM and ScaDMT, both small membrane proteins of ∼50 kDa. Using an additional anti-Fab nanobody further facilitated reliable initial three-dimensional structure determination from small cryo-EM test datasets. Given that NabFab is of synthetic origin, is humanized, and can be conveniently expressed in Escherichia coli in large amounts, it may be useful not only for structural biology but also for biomedical applications.

Synthesis of a Poly(3-dodecylthiophene) Bearing Aniline Groups for the Covalent Functionalization of Carbon Nanotubes

The functionalization of carbon nanotubes by polymers necessitates two steps, first their modification by oxidizing them or by covalently attaching small compounds to them, then the growth of the polymer chains from these anchors or their grafting onto them. In order to better control the process and the rate of functionalization, we develop polymers able to covalently react with the carbon nanotubes by their side chains in one step. We describe the synthesis of a copolymer of dodecylthiophene and its analogue bearing an aniline group at the end of the dodecyl side chain. This copolymer can functionalize single-walled carbon nanotubes (SWNTs) non-covalently and disperse more SWNTs than its hexyl analogues. UV-Vis and fluorescence spectroscopies show that in these non-covalent hybrids, the polymer forms p-stacked aggregates on the SWNTs. The non-covalent hybrids can be transformed into covalent ones by diazonium coupling. In these covalent hybrids the polymer is no longer p-stacked. According to Raman spectroscopy, the conformation of the poly(3-hexylthiophene) backbone is more ordered in the non-covalent hybrids than in the covalent ones.

Antibody Humanization: Intra and Inter VH-VL Non-bond Energy Sort Best Humanized Antibody

Antibody humanization of non-human derived antibody reduces immunogenicity of antibody drug. Computer aided antibody humanization has became an efficient and rapid routine process. Our computational humanization pipeline includes CDR grafting onto antibody crystal structure and humanization of CDR grafted antibody structure. Then, intra and inter VH-VL non-bond energy is calculated and sorted for selection best humanized antibody. Compare to experimental dataset, result indicate that intra and inter VH-VL non-bond energy could rank humanized antibody.