Multiple Gene Expression in Cell-Free Protein Synthesis Systems for Reconstructing Bacteriophages and Metabolic Pathways.

As a fast and reliable technology with applications in diverse biological studies, cell-free protein synthesis has become popular in recent decades. The cell-free protein synthesis system can be considered a complex chemical reaction system that is also open to exogenous manipulation, including that which could otherwise potentially harm the cell's viability. On the other hand, since the technology depends on the cell lysates by which genetic information is transformed into active proteins, the whole system resembles the cell to some extent. These features make cell-free protein synthesis a valuable addition to synthetic biology technologies, expediting the design-build-test-learn cycle of synthetic biology routines. While the system has traditionally been used to synthesize one protein product from one gene addition, recent studies have employed multiple gene products in order to, for example, develop novel bacteriophages, viral particles, or synthetic metabolisms. Thus, we would like to review recent advancements in applying cell-free protein synthesis technology to synthetic biology, with an emphasis on multiple gene expressions.

Molecular identification of ascidians from the Palk Bay Region, Southeast coast of India.

BACKGROUND: More than 50 ascidian species distributed in the Palk Bay, Southeast coat of India. Up to a very few molecular work has been performed to determine the evolutionary relationships of ascidians in the world. OBJECTIVES: Present study explored the value of mtDNA data in assessing phylogenetic relationships within the family Ascidiacea, Didemnidae, Styelidae, and molecular identification of ascidians from the Palk Bay, Southeast coast of India. MATERIALS AND METHODS: The phylogeny analysis was executed based on mitochondrial cytochrome oxidase subunit I (COI) sequences of ascidian species. A BLASTN search can be run to determine the similarity of an unknown DNA sequence (query) with the collection of all known DNA sequences in GenBank. RESULT: The BLASTN results showed that Didemnum candidum, Ascidia ahodori and Styela clava match the other listings for these species in GenBank. Mitochondrial COI gene sequences of collected ascidians were submitted to GenBank and obtained the accession numbers. CONCLUSION: This study is the first report of molecular identification of ascidian species of the Palk Bay, Southeast coast of India. This information about Palk Bay ascidian communities provides a baseline of general biodiversity of that ecosystem.

A statistical approach for optimization of alkaline lipase production by ascidian associated-Halobacillus trueperi RSK CAS9.

A marine ascidian-associated bacterium, Halobacillus trueperi RSK CAS9, was optimized for lipase production by response surface methodology using marine waste as substrate. The central composite design was employed, and the optimal medium constituents for maximum lipase production (1355.81 U/ml) were determined to be tuna powder (14.58 g/l), olive oil (5.05 ml/l); NaCl (72.42 g/l), temperature (45 °C) and pH 9.0. An alkaline lipase was purified to 8.46 fold with 1193.59 U mg-1 specific activities with the molecular weight of 44 kDa. The activity was substantially inhibited by EDTA and PMSF, indicating that it was a metalloenzyme serine residue which was essential for catalytic activity. Thus, lipase production by microbial conversion of marine fish wastes in this study suggested its potential utilization for the production of high value products.