scholarly journals Transcriptome Analysis Reveals the Procyanidin Treatment-responsive Genes Involved in Regulating Procyanidin Accumulation during Banana Ripening and Senescence

2021 ◽  
pp. 1-9
Author(s):  
Chao Zhou ◽  
Haide Zhang ◽  
Yixing Li ◽  
Fenfang Li ◽  
Jiao Chen ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Transcriptome Analysis ◽  
Musa Acuminata ◽  
Specific Gene ◽  
Biosynthetic Enzymes ◽  
Chain Reaction ◽  
Leucoanthocyanidin Reductase ◽  
Gene Functions ◽  
Polymerase Chain ◽  
Key Genes

The mechanism regulating procyanidin (PA) accumulation in banana (Musa acuminata) fruit is not understood. During this study, the effects of PA treatment on the activities of banana PA biosynthetic enzymes and transcriptomic profiles were investigated. The results showed that PA treatment delayed the decreases in leucoanthocyanidin reductase and anthocyanidin reductase activities, which affected the accumulation of PA. Furthermore, the peel samples of the control fruit and the PA-treated fruit on day 1 were selected for transcriptomic analysis. The results revealed that PA treatment induced 1086 differentially expressed genes. Twenty-one key genes, including those encoding biosynthetic enzymes and regulatory factors involved in PA biosynthesis, were validated using a quantitative real-time polymerase chain reaction. The results showed that these genes were upregulated by PA treatment during banana storage. Taken together, our study improves current understanding of the mechanism underlying PA-regulated banana senescence and provide new clues for investigating specific gene functions.

Detection of specific mRNAs in single nephron segments by use of the polymerase chain reaction

1990 ◽  
Vol 258 (5) ◽  
pp. F1470-F1474 ◽  
Author(s):  
T. Moriyama ◽  
H. R. Murphy ◽  
B. M. Martin ◽  
A. Garcia-Perez
Keyword(s):  
Polymerase Chain Reaction ◽  
Aldose Reductase ◽  
Collecting Duct ◽  
Specific Gene ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Nephron Segments ◽  
Single Nephron ◽  
Specific Mrnas ◽  
Polymerase Chain

We have developed a procedure to detect specific mRNAs in single renal nephron segments. This approach combines microdissection, reverse transcription (RT) of the target mRNA, and amplification of the resulting cDNA using the polymerase chain reaction (PCR). After microdissection, the sample is placed in a tube where it is permeabilized and where all reactions are performed directly without the need for isolation of the RNA. Our model target was the mRNA for aldose reductase. This enzyme catalyzes the conversion of glucose to sorbitol. Its expression is modulated by changes in extracellular osmolality in the renal medulla. RT-PCR of inner medullary collecting duct (1 mm) and glomeruli (6-10) yielded a product of the predicted length (670 base pairs) defined by the PCR primers. Its identity was confirmed by a specific oligonucleotide probe that differed from the primers. RT-PCR of proximal tubules (1 mm) resulted in no aldose reductase-specific amplification product. RT-PCR is generally applicable for measuring specific gene expression in single nephron segments or small numbers of cultured cells. Utility, limitations, and refinements of this approach are discussed.

scholarly journals Investigation of Different Commercially Available Real Time-Polymerase Chain Reaction Kits for SARS-CoV-2 Diagnosis

2021 ◽  
Author(s):  
Rajeev Kumar Jain ◽  
Nagaraj Perumal ◽  
Rakesh Shrivastava ◽  
Kamlesh Kumar Ahirwar ◽  
Jaya Lalwani ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Internal Control ◽  
Housekeeping Genes ◽  
Housekeeping Gene ◽  
Specific Gene ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Gene Target ◽  
Polymerase Chain

Introduction: The whole world is facing an ongoing global health emergency of COVID-19 disease caused by the Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). Real-Time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) is a gold standard in the detection of SARS-CoV-2 infection. Presently, many single tube multiple gene target RT-PCR kits have been developed and are commercially available for Coronavirus Disease 2019 (COVID-19) diagnosis. Aim: To evaluate the performance of seven COVID-19 RT-PCR kits (DiagSure, Meril, VIRALDTECT II, TruPCR, Q-line, Allplex and TaqPath) which are commercially available for COVID-19 RT-PCR diagnosis. Materials and Methods: This observational study was conductedat the State Virology Laboratory (SVL), Gandhi Medical College, Bhopal, Madhya Pradesh, India. Seven commercially available kits have been evaluated on the basis of: (i) number of SARS-CoV-2 specific gene target; (ii) human housekeeping genes as internal control; (iii) RT-PCR run time; and (iv) kit performances to correctly detect SARS-CoV-2 positive and negative RNA samples. A total of 50 RNA samples (left over RNA) were included, master mix preparation, template addition and RT-PCR test has been performed according to kits literature. At the end of PCR run, mean and standard deviation of obtained cut-off of all kits were calculated using Microsoft Excel. Results: All seven RT-PCR kits performed satisfactory regarding the reproducibility and they could correctly identify 30 positive and 20 negative RNA samples. RNA samples (group C) having low viral loads with a high Cycle threshold (Ct) value (>30) were also detected by all these seven kits. Obtained Ct values of each group was in parallel range in comparison with the initial testing Ct values. Kits were found to be superior which contains primers and probes for three SARS-CoV-2 specific gene targets, have human housekeeping gene as internal control and taking less time to complete RT-PCR. Conclusion: All seven COVID-19 RT-PCR kits included in this study demonstrated satisfactory performance and can be used for the routine molecular diagnosis of COVID-19 disease.

scholarly journals Rapid Real-Time Polymerase Chain Reaction for Salmonella Serotyping Based on Novel Unique Gene Markers by Pangenome Analysis

2021 ◽  
Vol 12 ◽  
Author(s):  
Seung-Min Yang ◽  
Eiseul Kim ◽  
Dayoung Kim ◽  
Hyeon-Be Kim ◽  
Jiwon Baek ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Real Time Pcr ◽  
Pathogenic Bacteria ◽  
Specific Gene ◽  
Chain Reaction ◽  
Gene Markers ◽  
Unique Gene ◽  
High Prevalence ◽  
Polymerase Chain

An accurate diagnostic method for Salmonella serovars is fundamental to preventing the spread of associated diseases. A diagnostic polymerase chain reaction (PCR)-based method has proven to be an effective tool for detecting pathogenic bacteria. However, the gene markers currently used in real-time PCR to detect Salmonella serovars have low specificity and are developed for only a few serovars. Therefore, in this study, we explored the novel unique gene markers for 60 serovars that share similar antigenic formulas and show high prevalence using pangenome analysis and developed a real-time PCR to detect them. Before exploring gene markers, the 535 Salmonella genomes were evaluated, and some genomes had serovars different from the designated serovar information. Based on these analyses, serovar-specific gene markers were explored. These markers were identified as genes present in all strains of target serovar genomes but absent in strains of other serovar genomes. Serovar-specific primer pairs were designed from the gene markers, and a real-time PCR method that can distinguish between 60 of the most common Salmonella serovars in a single 96-well plate assay was developed. As a result, real-time PCR showed 100% specificity for 199 Salmonella and 29 non-Salmonella strains. Subsequently, the method developed was applied successfully to both strains with identified serovars and an unknown strain, demonstrating that real-time PCR can accurately detect serovars of strains compared with traditional serotyping methods, such as antisera agglutination. Therefore, our method enables rapid and economical Salmonella serotyping compared with the traditional serotyping method.

Sign in / Sign up

Export Citation Format

Share Document