Authors:Anna Tóth, Katalin Fodor, P. Blazsó, I. Cserpán, Tünde Praznovszky, V. Tubak, A. Udvardy, Gy. Hadlaczky, and R. Katona
Direct reprogramming of mouse fibroblasts into induced pluripotent stem cells (iPS) was achieved recently by overexpression of four transcription factors encoded by retroviral vectors. Most of the virus vectors, however, may cause insertional mutagenesis in the host genome and may also induce tumor formation. Therefore, it is very important to discover novel and safer, non-viral reprogramming methods. Here we describe the reprogramming of somatic cells into iPS cells by a novel protein-based technique. Engineered Oct4, Sox2 and Klf4 transcription factors carrying an N-terminal Flag-tag and a C-terminal polyarginine tail were synthesized by a recently described mammalian artificial chromosome expression system (ACEs). This system is suitable for the high-level production of recombinant proteins in mammalian tissue culture cells. Recombinant proteins produced in this system contain all the post-translational modifications essential for the stability and the authentic function of the proteins. The engineered Oct4, Sox2 and Klf4 proteins efficiently induced the reprogramming of mouse embryonic fibroblasts by means of protein transduction. This novel method allows for the generation of iPS cells, which may be suitable for therapeutic applications in the future.
Authors:Theodora Tsiligianni, Irene Valasi, Sándor Cseh, Emmanuel Vainas, Vera Faigl, Fotini Samartzi, Thomas Papanikolaou, Eleni Dovolou, and Georgios Amiridis
Follicular development and oocyte quality were assessed by laparoscopic observation and
fertilisation, respectively, in melatonin-treated (Group M) and control (Group C) anoestrous Chios ewes (n = 10 in each group). Fourteen days after melatonin insertion, all ewes had laparoscopic evaluation of the follicular population followed by oocyte pick-up (OPU); on day 22 intravaginal progestagen sponges were inserted for 14 days. Two days after sponge removal the follicular population was re-evaluated and a second follicular aspiration was performed. Collected oocytes from the second OPU underwent
maturation, fertilisation and culture. The number of large follicles was higher in Group M than in the control ewes during the first OPU and tended to be so (P = 0.06) at the second. Morphologically, oocytes collected from controls were of better quality than those from Group M; however, more oocytes collected from melatonintreated animals fertilised and developed
. These results indicate that melatonin is a potent regulator of follicular development and oocyte competence during the anoestrous period of the ewe.
Authors:Krisztián Bányai, Jelle Matthijnssens, György Szücs, Petra Forgách, Károly Erdélyi, Marc van Ranst, Eleonora Lorusso, Nicola Decaro, Gabriella Elia, and Vito Martella
In rotaviruses, intragenic recombination or gene rearrangement occurs almost exclusively in the genome segments encoding for non-structural proteins. Rearranged RNA originates by mechanisms of partial sequence duplications and deletions or insertions of non-templated nucleotides. Of interest, epidemiological investigations have pointed out an unusual bias to rearrangements in genome segment 11, notably in rotavirus strains of lapine origin, as evidenced by the detection of numerous lapine strains with super-short genomic electropherotype. The sequence of the full-length genome segment 11 of two lapine strains with super-short electropherotype, LRV-4 and 3489/3, was determined and compared with rearranged and normal cognate genome segments of lapine rotaviruses. The rearranged genome segments contained head-to-tail partial duplications at the 3′ end of the main ORF encoding NSP5. Unlike the strains Alabama and B4106, intermingled stretches of non-templated sequences were not present in the accessory RNA of LRV-4 and 3489/3, while multiple deletions were mapped, suggesting the lack of functional constraints. Altogether, these findings suggest that independent rearrangement events have given origin to the various lapine strains that have super-short genome pattern.
Authors:Y. Yang, X. Chen, Z. He, M. Röder, and L. Xia
Genotypes with various
alleles perform different levels of pre-harvest sprouting (PHS) tolerance. In this study, 217 white-grained wheat cultivars, including 75 landraces, 39 historical cultivars, and 103 modern cultivars from five major regions of China, were examined to characterize the diversity of the Viviparous-1B (
) locus associated with PHS tolerance. Four
alleles were identified, three (
) of which were previously reported in Chinese wheat cultivars. A new allele,
, was identified in the PHS tolerant landrace Hongheshangtou. Sequence analysis showed that
had an insertion of a 4-bp fragment, two SNPs, and a deletion of an 83-bp fragment compared with the nucleotide sequence of
(AJ400713), all located in the third intron.
shared 97.80% similarity with the nucleotide sequence of AJ400713. The frequencies of
were 36.0%, 5.3%, and 57.3% in landraces; 23.1%, 7.7%, and 69.2% in historical cultivars; and 52.4%, 0%, and 47.6% in current cultivars, respectively.
Authors:J. González, P. Rubio, A. López, and N. Jouve
androgenesis allows haploid and double haploid homozygous plants to be obtained. However, in cereals, non-viable albino plants are often produced. This limits the use of this technique both in basic studies and in plant genetic improvement programs. We have analyzed eight microsatellites of the chloroplast genome (cpSSR) in a collection of 46 double haploid green lines derived from the intervarietal hybrid ‘Torote’ × ‘Presto’, and 49 haploid albino plants all obtained from the double haploid line ‘ATOPE-41’, in order to know the stability of this sequences after the androgenetic process. The green plants showed no variations at the analyzed loci, while two of the albino plants differed at two loci: WCt2 and WCt9. These variations notably affected the microsatellite nucleotide sequences as well as the sequences surrounding them. The changes observed consisted of insertions, deletions and nucleotide substitutions, with transition and transversion types equally common. The latter albino plants showed several alleles at the polymorphic loci, indicating the existence of heteroplasmy.
Translating research articles is common practice, although relatively little data exists on the problems in this type of translation. This study examines the translation of textual metadiscourse in academic writing, using the example of translating Slovene research articles into English. The main part of the analysis examines textual metadiscourse in 30 geography articles in Slovene and their translations into English. In addition, the same investigation is conducted in a corpus of 30 English-original geography articles. The translation strategies are analysed at two levels. At the first level, the overall strategy is analysed: the metadiscourse items are compared in the originals and the translations and instances of matching expressions, omissions, and insertions are identified. At the second level, the corresponding expressions are compared and changes made in the translation are noted. The overall results are then compared to those of the comparable target language subcorpus. The results show that not all metadiscourse items found in the original texts are translated, while, at the same time, a significant number of items are inserted in the translation. For those metadiscourse items which are translated, literal translation is chosen in over half of the cases. The results for the comparable target language corpus reveal that metadiscourse is used more frequently in English originals than in translations from Slovene.
Authors:L. Wei, S.G. Bai, X.J. Hou, J.M. Li, B. Zhang, W.J. Chen, D.C. Liu, B.L. Liu, and H.G. Zhang
Among 20 awnless Tibetan wheat cultivars analyzed by SDS-PAGE, the migration rate of an HMW-GS in XM001584 and XM001593, named 1BX23*. was shown to be slightly faster than 1Bx6. and slower than Bx7. Its nucleotide sequence was isolated based on homology clones. In a phylogenetic tree of 1Bx genes, 1Bx23* was apparently clustered with 1Bx23. Compared with 1Bx23. eight single nucleotide replacements caused four single amino acid replacements in 1Bx23*. The deletion of “G” at base pair 1463 and insertion of “A” at 1509 bps induced a 42-nucleotide frame shift. “GQRQQAGQWQRPGQ” was replaced by “DKGNRQDNGNDRDK”. The new segment cannot be found in other HMW-GSs, and it is very similar to a segment found in collagen. Moreover, an 18-nucleotide deletion made 1Bx23* six amino acids shorter than 1Bx23. The cultivar XM001593 had 28 chromosomes, which signifies that it was tetraploid wheat, and that the new HMW-GS 1Bx23* cannot be used directly for breeding in common wheat.
Authors:Z.L. Li, H.Y. Li, G. Chen, X.J. Liu, C.L. Kou, S.Z. Ning, Z.W. Yuan, M. Hao, D.C. Liu, and L.Q. Zhang
Seven Glu-A1m allelic variants of the Glu-A1mx genes in Triticum monococcum ssp. monococcum, designated as 1Ax2.1a, 1Ax2.1b, 1Ax2.1c, 1Ax2.1d, 1Ax2.1e, 1Ax2.1f, and 1Ax2.1g were characterized. Their authenticity was confirmed by successful expression of the coding regions in E. coli, and except for the 1Ax2.1a with the presence of internal stop codons at position of 313 aa, all correspond to the subunit in seeds. However, all the active six genes had a same DNA size although their encoding subunits showed different molecular weight. Our study indicated that amino acid residue substitutions rather than previously frequently reported insertions/deletions played an important role on the subunit evolution of these Glu-A1mx alleles. Since variation in the Glu-A1x locus in common wheat is rare, these novel genes at the Glu-A1mx can be used as candidate genes for further wheat quality improvement.
High-molecular-weight glutenin subunits (HMW-GSs) are important seed storage proteins associated with bread-making quality in common wheat (Triticum aestivum L., 2n = 6x = 42, AABBDD). Variation in the Glu-A1x locus in common wheat is scare. Diploid Triticum monococcum ssp. monococcum (2n = 2x = 14, AmAm) is the first cultivated wheat. In the present study, allelic variations at the Glu-A1mx locus were systematically investigated in 197 T. monococcum ssp. monococcum accessions. Out of the 8 detected Glu-A1mx alleles, 5 were novel, including Glu-A1m-b, Glu-A1m-c, Glu-A1m-d, Glu-A1m-g, and Glu-A1m-h. This diversity is higher than that of common wheat. Compared with 1Ax1 and 1Ax2*, which are present in common wheat, these alleles contained three deletions/insertions as well as some single nucleotide polymorphism variations that might affect the elastic properties of wheat flour. New variations in T. monococcum probably occurred after the divergence between A and Am and are excluded in common wheat populations. These allelic variations could be used as novel resources to further improve wheat quality.
Despite intensive efforts in recent decades to develop preventive or therapeutic vaccines against diseases caused by herpes simplex virus (HSV), or varicella-zoster virus (VZV), members of the Alpha herpes virinae subfamily of human herpes viruses,a safe and efficient vaccine has been approved for commercial development only against VZV. The VZV vaccine contains a live attenuated strain, OKA. It consists of amixture of at least 13 subpopulations of viruses, all with deletions, insertions or mutations in the genome; the most common mutations are observed in the open reading frame 62 (ORF62). Experience over more than 30 years in Japan, the USA and other countries where VZV vaccination is provided has demonstrated that the vaccine is safe and the effectiveness of two doses compared to unvaccinated children is 98–99%. When administered in a higher dose to stimulate the declining cell-mediated immunity, the same vaccine has been shown to reduce the incidence and severity of herpes zoster in immunocompetent individuals older than 60 years. Vaccination of immuno-compromised subjects with this VZV vaccine is problematic and various strategies need to be explored. Differences in the pathomechanisms of infection, latency and immune evasion of VZV and HSV, together with host genetic factors, may explain the availability of the successful VZV vaccine and the failures of the past HSV vaccine candidates.