Aim: To study the effect of integration of tandem aroG-pheA genes into the tyrA locus of Corynebacterium glutamicum (C. glutamicum) on the production of L-phenylalanine.
Methods: By nitrosoguanidine mutagenesis, five p-fluorophenylalanine (FP)-resistant mutants of C.glutamicum FP were selected. The tyrA gene encoding prephenate dehydrogenase (PDH) of C.glutamicum was amplified by polymerase chain reaction (PCR) and cloned on the plasmid pPR. Kanamycin resistance gene (Km) and the P(BF) -aroG-pheA-T (GA) fragment of pGA were inserted into tyrA gene to form targeting vectors pTK and pTGAK, respectively. Then, they were transformed into C.glutamicum FP respectively by electroporation. Cultures were screened by a medium containing kanamycin and detected by PCR and phenotype analysis. The transformed strains were used for L-phenylalanine fermentation and enzyme assays.
Results: Engineering strains of C.glutamicum (Tyr(-)) were obtained. Compared with the original strain, the transformed strain C. glutamicum GAK was observed to have the highest elevation of L-phenylalanine production by a 1.71-fold, and 2.9-, 3.36-, and 3.0-fold in enzyme activities of chorismate mutase, prephenate dehydratase and 3-deoxy-D-arabinoheptulosonate-7-phosphate synthase, respectively.
Conclusion: Integration of tandem aroG-pheA genes into tyrA locus of C. glutamicum chromosome can disrupt tyrA gene and increase the yield of L-phenylalanine production.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4612019 | PMC |
| http://dx.doi.org/10.3748/wjg.v10.i24.3683 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
T-cell receptor beta variable gene polymorphism predicts immune-related adverse events during checkpoint blockade immunotherapy.
J Immunother Cancer
August 2023
Investigational Cancer Therapeutics, University of Texas MD Anderson Cancer Center, Houston, Texas, USA
Background: Immune checkpoint inhibitors have revolutionized cancer treatment. However, they are associated with a unique spectrum of side effects, called immune-related adverse events (irAEs), which can cause significant morbidity and quickly progress to severe or life-threatening events if not treated promptly. Identifying predictive biomarkers for irAEs before immunotherapy initiation is therefore a critical area of research.
View Article and Find Full Text PDFVariations in Fibrinogen-like 1 () Gene Locus as a Genetic Marker Related to Fat Deposition Based on Pig Model and Liver RNA-Seq Data.
Genes (Basel)
August 2022
Department of Pig Breeding, National Research Institute of Animal Production, Krakowska 1, 32-083 Balice, Poland.
The goal of this study was to evaluate the effects of mutations in the gene associated with pig productive traits to enrich the genetic marker pool for further selection and to support the studies on in the context of the fat deposition (FD) process. The variant calling and χ analyses of liver RNA-seq data were used to indicate genetic markers. mutations were genotyped in the Złotnicka White (n = 72), Polish Large White (n = 208), Duroc (n = 72), Polish Landrace (PL) (n = 292), and Puławska (n = 178) pig breeds.
View Article and Find Full Text PDFHaplotype-resolved germline and somatic alterations in renal medullary carcinomas.
Genome Med
July 2021
Department of Pediatrics, Emory University, Atlanta, GA, USA.
Background: Renal medullary carcinomas (RMCs) are rare kidney cancers that occur in adolescents and young adults of African ancestry. Although RMC is associated with the sickle cell trait and somatic loss of the tumor suppressor, SMARCB1, the ancestral origins of RMC remain unknown. Further, characterization of structural variants (SVs) involving SMARCB1 in RMC remains limited.
View Article and Find Full Text PDFA CRISPR Competition Assay to Identify Cancer Genetic Dependencies.
Bio Protoc
July 2020
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
The CRISPR/Cas9 system is a powerful tool for genome editing, wherein the RNA-guided nuclease Cas9 can be directed to introduce double-stranded breaks (DSBs) at a targeted locus. In mammalian cells, these DSBs are typically repaired through error-prone processes, resulting in insertions or deletions (indels) at the targeted locus. Researchers can use these Cas9-mediated lesions to probe the consequences of loss-of-function perturbations in genes of interest.
View Article and Find Full Text PDFAssociation between single nucleotide polymorphisms in GPAT1 locus and pork quality in pigs.
Meat Sci
April 2020
Department of Pig Breeding, National Research Institute of Animal Production, Poland, 1 Krakowska, 32-083 Balice, Poland.
GPAT1 gene is considered to be a genetic marker for intramuscular fat content. The GPAT enzymes catalyze the first step in triacylglycerol synthesis. In the present study, the search for polymorphisms within the pig GPAT1 gene locus as well as association analyses between SNPs and meat quality traits were performed.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!