ABSTRACT
Adenovirus vectors possess a good safety profile, an extensive genome, a range of host cells, high viral yield, and the ability to elicit broad humoral and cellular immune responses. Adenovirus vectors are widely used in infectious disease research for future vaccine development and gene therapy. In this study, we obtained a fowl adenovirus serotype 4 (FAdV-4) isolate from sick chickens with hepatitis-hydropericardium syndrome (HHS) and conducted animal regression text to clarify biological pathology. We amplified the transfer vector and extracted viral genomic DNA from infected LMH cells, then recombined the mixtures via the Gibson assembly method in vitro and electroporated them into EZ10 competent cells to construct the FAdV-4 infectious clone. The infectious clones were successfully rescued in LMH cells within 15 days of transfection. The typical cytopathic effect (CPE) and propagation titer of FAdV-4 infectious clones were also similar to those for wild-type FAdV-4. To further construct the single-cycle adenovirus (SC-Ad) vector, we constructed SC-Ad vectors by deleting the gene for IIIa capsid cement protein. The FAdV4 infectious clone vector was introduced into the ccdB cm expression cassette to replace the IIIa gene using a λ-red homologous recombination technique, and then the ccdB cm expression cassette was excised by PmeI digestion and self-ligation to obtain the resulting plasmids as SC-Ad vectors.
Subject(s)
Communicable Diseases , Hepatitis A , Animals , Chickens , Serogroup , Adenoviridae/genetics , Capsid Proteins , DNA, ViralABSTRACT
The acoustics of lexical tones are highly variable across talkers, and require second-language (L2) learners' flexibility in accommodating talker-specific tonal variations for successful learning. This study investigated how tone training with high vs. low talker-variability modulated novice learners' neural responses to non-native tones. A passive oddball paradigm tested Mandarin-speaking participants' neural responses to Cantonese low-high and low-mid tonal contrasts in the pretest and posttest. Participants were trained using a tone identification task with feedback, either with high or low talker-variability. The results of mismatch negativity (MMN) showed no group difference in the pretest whereas the high-variability group demonstrated greater neural sensitivity to the low-high tonal contrast produced by a novel talker and a trained talker in the posttest. The finding provides (tentative) novel evidence that training variability may benefit perceptual learning of the relatively easy tone pair and facilitate the formation of talker-independent representations of non-native tones by novice learners.