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Cloning - Springer Nature

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Last Updated: 19 June 2022

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Reference-aided full-length transcript assembly, cDNA cloning, and molecular characterization of coronatine-insensitive 1b (COI1b) gene in coconut (Cocos nucifera L.)

"Background In the Philippines, 26% of the total agricultural land is devoted to coconut cultivation, making coconut is the country's most popular industrial crop. The coconut industry in the country's multimillion-dollar coconut industry is being threatened by the outbreak of coconut scale insect and other re-emerging insect pests that have urged national research institutes to work together on finding new tolerant coconut varieties. CnCOI1b-1 was 7919 bp with an ORF of 1176 amino acids decoded, while CnCOI1b-1 had a full-length cDNA with an ORF of 1743 bp encoding a deduced protein of 580 amino acids. Following the introduction of CSI relative to base levels, a seven-fold increase in COI1 gene expression in coconut was revealed by differential gene expression using qRT-PCR results. The findings of this research were also critical in the development of improved insect-resistant coconut varieties for vibrant coconut industry.

Source link: https://doi.org/10.1007/s11033-022-07658-w


Standardization of the PCR Technique for the Detection of the Toxoplasma gondii B1 Gene in Meat and Water Samples and Cloning of the Product for Use as Control

The parasite Toxoplasmosis is responsible for Toxoplasmosis in the parasite Toxoplasma gondii. The aim of this research was to standardize the PCR for the detection of the T. gondii B1 gene in meat and water samples, as well as cloning of the product for use as a benchmark. T. gondii DNA was found in 90% of the positive samples of meat and water with standardized PCR and using the cloned DNA as a control, but there was no amplification in the negative samples. Conclusions: T. gondii DNA testing in meat and water samples was found to be a quick way to determine T. gondii DNA in meat and water samples. This cloning of PCR product and its use as a marker of molecular diagnosis of toxoplasmosis may contribute to improved reproducibility of this technique and avoid the use of patient samples or cultures, which have numerous drawbacks.

Source link: https://doi.org/10.1007/s11686-022-00579-5


Molecular cloning and characterization of the wheat (Triticum aestivum L.) starch synthase III gene sheds light on its structure

"Using overlapping gene-specific primers and cloning the gene was used for PCR amplification of TaSS-IIIa1D gene and cloning of the TaSS-IIIa1D gene in the pJET1. 2/blunt cloning vector, as well as sequencing and measurement of SNPs was performed. " According to In-silico analysis, the TaSS-III gene contains two homologs, TaSS-IIIa and TaSS-IIIb, which were discovered on the plus strand of chromosome 1 and minus strands of chromosome 2, respectively. SNPs corresponding to the Chinese spring cultivar were identified in PBW343 and twenty in IC252874 genotypes for the gene vis-a-vis with the reference copy of the Chinese spring cultivar. Transition bias was also evident among the two genotypes for the gene consisting of 11 transitions and seven transversions. After proper validation, wheat improvement could be used for wheat improvement after proper validation. ".

Source link: https://doi.org/10.1007/s42976-021-00182-w


Cloning and homologous characterization of geranylgeranyl pyrophosphate synthase (GGPPS) from Withania somnifera revealed alterations in metabolic flux towards gibberellic acid biosynthesis

"Main conclusion Overexpression of a novel geranyl pyrophosphate synthase gene in planta resulted in increased levels of gibberellic acid and a decrease in withanolide content. " Abstract Due to its unique stockpile of pharmaceutically active secondary metabolites, Withania somnifera Dunal, the herb from family Solanaceae, is one of the most popular medicinal plant used in traditional therapeutic systems. In this research, a key enzyme in the terpenoid biosynthetic pathway viz. The complete WsGGPPS gene encoded a polypeptide of 365 amino acids, with 1,104 base pairs coding a polypeptide of 365 amino acids. WsGGPPS transcripts were predominantly expressed in flower tissue, followed by berry tissue, according to a quantitative expression review. WsGGPPS had close traces to Solanum tuberosum and Solanum pennellii, according to Amino acid sequence alignment and phylogenetic studies. According to a WsGGPPS overexpression in planta for its functional analysis, the WsGGPPS was involved in gibberellic acid biosynthesis. ".

Source link: https://doi.org/10.1007/s00425-022-03912-4


Cloning of the broadly effective wheat leaf rust resistance gene Lr42 transferred from Aegilops tauschii

"The wheat wild cousin Aegilops tauschii had been used to transfer the Lr42 leaf rust resistance gene from wheat to bread wheat before," Aegilops tauschii was used to convert the Lr42 leaf rust resistance gene from wheat wheat. " Overexpression of a nucleotide-binding gene leucine-rich repeat gene AET1Gv20040300 in wheat is responsible for a lot of leaf rust in wheat, as well as a gene mutation that altered the resistance. Documenting its widespread use and effect on crop improvement, Lr42's Cloning of Lr42 produces diagnostic indicators and over 1000 CIMMYT wheat lines carrying Lr42. The Aegilops tauschii -derived leaf rust resistance gene Lr42 has been used widely for breeding resistance wheat cultivars, but the exact source is uncertain. ".

Source link: https://doi.org/10.1038/s41467-022-30784-9

* Please keep in mind that all text is summarized by machine, we do not bear any responsibility, and you should always check original source before taking any actions

* Please keep in mind that all text is summarized by machine, we do not bear any responsibility, and you should always check original source before taking any actions