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DNA Structure, Sequencing, Synthesis, and Modification: Making Biology Molecular
Published in Richard J. Sundberg, The Chemical Century, 2017
In the 1990s Craig Venterp and Mark Adams, working at the NIH Institute for Neurological Diseases and Stroke, developed a new sequencing approach that promised faster results. The method was called the whole genome shotgun (WGS) approach. One of the crucial features of Venter’s approach was use of expressed sequence tags, or EST for short. These are derived from the m-RNA corresponding to active protein production. The complementary DNA strand, called c-DNA can be cloned into bacterial DNA, and, when sequenced provides information on functional DNA, that is, genes. The concept was originally introduced by Paul Schimmel of MIT. It was further developed by Sydney Brenner at the MRC laboratory in Cambridge and has the advantage of focusing attention on expressed gene sequences, while avoiding the introns that do not contain coding information. The EST are only partial fragments of genes, however, and do not provide any direct information about the function of the gene. However, they can be identified if other information on the gene sequence is available. Using this approach, by 1992, Venter had sequenced parts of about 2500 human genes.
Genomic Approaches for Understanding Abiotic Stress Tolerance in Plants
Published in Hasanuzzaman Mirza, Nahar Kamrun, Fujita Masayuki, Oku Hirosuke, Tofazzal M. Islam, Approaches for Enhancing Abiotic Stress Tolerance in Plants, 2019
Richa Rai, Amit Kumar Rai, Madhoolika Agrawal
The catalogue pertaining to expressed genes of a particular species is investigated through expressed sequence tags (ESTs). Focusing mainly on functional studies, EST techniques are used which are fast and cost-effective in identifying genes (Iqbal et al., 2013). The availability of EST compilations and cDNA sequences of Arabidopsis and rice has encouraged large-scale compilations for other crops as well (Bevan et al., 1999; Tyagi et al., 2006). However, there are limited studies focusing on ESTs from plants exposed to abiotic stresses. The main importance of the EST technique is to recognize the vital function of the responsible genes. The National Centre for Biotechnological Information (NCBI) database currently has about a million ESTs for crops like maize, rice, soybean, and wheat, along with other plants as well. In order to develop plant EST datasets with respect to stress-responsive genes, it is necessary to build up sequencing programs at different developmental stages based on cDNA libraries from stress-treated plant tissues and organs of different plant species. In comparison to cDNA, ESTs are shorter, and their overlapping provides more information about the organization of parental cDNA. Since para-log genes may result in sequences being misassembled, they must be handled carefully, particularly in species with polyploidy (Rudd, 2003). They are widely employed in crops which comprise of lengthy and repetitive genomes. They have the potential to aid in gene discovery. The latest reports have shown the EST sequencing method as being most suitable for evaluating the range of genotypes under controlled and stressed conditions (AkpJnar et al., 2013; Brenner et al., 2000).
Renewable biofuels from microalgae: technical advances, limitations and economics
Published in Environmental Technology Reviews, 2023
Subhisha Raj, Amrutha Sajith, Arathi Sreenikethanam, Swathi Vadlamani, Aiswarya Satheesh, Anurup Ganguly, J. Rajesh Banu, Sunita Varjani, Poornachandar Gugulothu, Amit K. Bajhaiya
Genetic engineering is a promising method to enhance microalgal biomass and biofuel production. Availability of microalgal genome sequences will accelerate the progress of the genetic engineering in future. As microalgal growth depends upon various physical stress parameters, genetic engineering strategies for modifying certain genes to control these environmental stress conditions would benefit enhanced microalgal biomass production [119]. One of the strategies includes reducing the number of light-harvesting complexes (LHC) or reducing chlorophyll antenna size to minimize the light-absorbing capacity of microalgae [120]. Genetic engineering strategies like sequencing nuclear, mitochondrial, and chloroplast genomes and establishing expressed sequence tag (EST) databases paved the path for enhanced biofuel production from microalgae [121].