Modeling the use of durational information in human spoken-word recognition

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Efficient expression of the yeast metallothionein gene in Escherichia coli.

The yeast metallothionein gene CUP1 was cloned into a bacterial expression system to achieve efficient, controlled expression of the stable, unprocessed protein product. The Escherichia coli-synthesized yeast metallothionein bound copper, cadmium, and zinc, indicating that the protein was functional. Furthermore, E. coli cells expressing CUP1 acquired a new, inducible ability to selectively sequester heavy metal ions from the growth medium

High-level production of fully active human alpha 1-antitrypsin in Escherichia coli.

The human alpha-1-antitrypsin (A1AT) gene expressed in Escherichia coli as a full-length, non-fusion gene product accumulates to a relatively low level approaching less than or equal to 0.1% of total cellular protein. In contrast, deletion of the first 5, 10 or 15 codons leads to production of truncated A1AT derivatives at levels between 10 and 30% of total cellular protein. The protein with the largest truncation was insoluble and inactive following solubilization by chaotropic agents. In contrast, the two derivatives with the smaller truncations were found to be soluble, and exhibit identical specific activities in both trypsin and elastase inhibition assays to authentic human A1AT. The expression of the full-length A1AT was also optimized by making silent third position mutations within its first 15 codons. These mutations were chosen to optimize codon usage and minimize the possibility of RNA secondary structure formation in this region. Via this approach, expression of full-length, authentic, fully active A1AT was increased at least 20-fold to 2% of total cellular protein. Optimal expression was obtained using as few as three silent mutations in the first five codons, confirming the importance of this 5'-terminal region as had been defined by our deletion mutants. Both the full-length derivatives as well as the small N-terminal deletion derivative can be readily purified from bacterial extracts in fully active form suitable for the examination of their potential therapeutic application

The modulation of lexical competition by segment duration

Item does not contain fulltextIn an eye-tracking study, we examined how fine-grained phonetic detail, such as segment duration, influences the lexical competition process during spoken word recognition. Dutch listeners' eye movements to pictures of four objects were monitored as they heard sentences in which a stop-initial target word (e.g., pijp "pipe") was preceded by an [s]. The participants made more fixations to pictures of cluster-initial words (e.g., spijker "nail") when they heard a long [s] (mean duration, 103 msec) than when they heard a short [s] (mean duration, 73 msec). Conversely, the participants made more fixations to pictures of the stop-initial words when they heard a short [s] than when they heard a long [s]. Lexical competition between stop- and cluster-initial words, therefore, is modulated by segment duration differences of only 30 msec.6 p