Using shotgun sequence data to find active restriction enzyme genes
نویسندگان
چکیده
Whole genome shotgun sequence analysis has become the standard method for beginning to determine a genome sequence. The preparation of the shotgun sequence clones is, in fact, a biological experiment. It determines which segments of the genome can be cloned into Escherichia coli and which cannot. By analyzing the complete set of sequences from such an experiment, it is possible to identify genes lethal to E. coli. Among this set are genes encoding restriction enzymes which, when active in E. coli, lead to cell death by cleaving the E. coli genome at the restriction enzyme recognition sites. By analyzing shotgun sequence data sets we show that this is a reliable method to detect active restriction enzyme genes in newly sequenced genomes, thereby facilitating functional annotation. Active restriction enzyme genes have been identified, and their activity demonstrated biochemically, in the sequenced genomes of Methanocaldococcus jannaschii, Bacillus cereus ATCC 10987 and Methylococcus capsulatus.
منابع مشابه
DNA sequence analysis and the computer.
Analysis of DNA sequences has greatly aided our understanding of molecular biology and genetics. Nucleotide sequences have provided a wealth of information about gene structure and function. In addition to revealing the primary structure of genes, sequencing has uncovered the phenomena of overlapping genes, segmented genes, pseudogenes and a different genetic code for mitochondria. To date over...
متن کاملCLONING AND EXPRESSION OF LEISHMANOLYSIN GENE FROM LEISHMANIA MAJOR IN PRIMATE CELL LINES
Leishmanolysin is a worldwide disease that is caused by different species of the genus Leishmania. Leishmanolysin, One of the genes expressed by Leishmania, appears to be an ideal candidate for genetic vaccination. In this study, a full length sequence, which encodes Leishmanolysin functionally critical regions (amino acids 100-579), was cloned from a Leishmania strain endemic to Iran. Analysis...
متن کاملDetection of New Silent Mutation at 348 bp Position in a CD18 Gene in Holstein Cattle Normal and Heterozygous for Bovine Leukocyte Adhesion Deficiency Syndrome
In India, Holstein and its crosses are being used extensively in breeding programmes and all these breeding bulls are screened for autosomal recessive genes. Blood samples are collected in ethylenediaminetetraacetic acid (EDTA) coated tubes and DNA was isolated by using phenol-chloroform method. Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) wereperformed by using...
متن کاملConstruction of an Expression Vector Containing a Novel Fusion Sequence from Middle Region of NS3 and Truncated Core Genes of Hepatitis C Virus
Background and Aims: DNA constructs containing HCV antigens have become one of the vaccine candidates for induction of anti-HCV cellular and humoral immunity. In this study, we constructed a novel expressing vector harboring a fusion sequence derived from an overlapping fragment in the middle of NS3 and a truncated core fragment to avoid troubles reported to be associated with full gene express...
متن کاملThe modified recombinant proinsulin: a simple and efficient route to produce insulin glargine in E. coli
Background: Recombinant insulin glargine, a long-acting analogue of insulin, is expressed as proinsulin in host cell and after purification and refolding steps cleaved to active insulin by enzymatic digestion using trypsin and carboxypeptidase B. Since the proinsulin's B and C chains have several internal arginine and lysine residues, a number of impurities are generated following treatment wit...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره 37 شماره
صفحات -
تاریخ انتشار 2009