A Non-toxic and Efficient Method for Extracting DNA and RNA from Peanut

[Objectives]To establish a non-toxic and efficient method for extracting DNA and total RNA from peanuts and laying a solid foundation for the molecular biology study of peanuts.[Methods]Based on the principle and method of purifying nucleic acids by silica gel adsorption at high salt and low pH condition,a non-toxic and efficient method to extract peanut DNA and total RNA using cetyltrimethyl ammonium bromide(CTAB)extraction solution was designed.The quality and purity of nucleic acids were detected by agarose gel electrophoresis and nucleic acids protein analyzer,respectively.The quality of DNA was further verified by enzyme digestion and PCR amplification using molecular marker techniques.The quality of total RNA was further verified by reverse transcription(RT)-PCR of actin gene and cDNA-SCoT gene differential display technique.[Results]The agarose gel electrophoresis test showed that the peanut DNA extracted by a low-toxic and effective method is free of contamination and degradation.Through the detection by the nucleic acid protein analyzer,the DNA concentration,yield,A260/A280 and A260/A230 of 5 peanut varieties were 419.6-498.2 ng/μL,20.98-24.91μg/g,1.89-1.96 and 2.03-2.28,respectively.The DNA was of high quality and can be completely digested by EcoRI restriction enzymes,and also can be used for SCoT and SRAP molecular marker technology analysis.The RNA extracted from different tissues of peanuts showed no visible DNA bands by non-denaturing agarose gel electrophoresis.The separated 28S bands were brighter than 18S.The ratio of A260/A280 and A260/A230 showed that the RNA quality was good and can be used for reverse transcription,RT-PCR of actin gene and amplification of cDNA-SCoT gene differential display technique.[Conclusions]This experiment established a low-toxic and effective method for extracting DNA and total RNA from peanuts.Compared with traditional methods,this method is more time-saving and cheaper than commercial kits.The most important point is that this method does not use toxic reagents such as phenol,chloroform and isopropanol.Thus,it is expected to be widely applied in molecular biology research.

Convenient synthesis of nucleoside 5'-(α-P-thio)triphosphates and phosphorothioate nucleic acids(DNA and RNA)

Modified deoxy-and ribo-nucleoside triphosphates are chemically synthesized in multiple steps due to the protection and deprotection of the nucleoside functionalities.To conveniently synthesize the S-modified triphosphates for enzymatically preparing phosphorothioate DNAs and RNAs(PS-DNA and PS-RNA) as potential therapeutics,herein we report a one-pot strategy to synthesize the deoxy-and ribo-nucleoside 5'-(α-P-thio)triphosphates(dNTPαS and NTPαS) without protecting any nucleoside functionalities.This facile synthesis is achieved by treating the nucleosides with a mild phosphitylating reagent,reacting selectively with the 5'-hydroxyl group of each unprotected nucleoside,followed by sulfurization and hydrolysis to afford the crude dNTPαS and NTPαS analogs(mixtures of Sp and Rp diastereomers).We also demonstrated that after just simple precipitation(without HPLC and ion-exchange purification),the quality of the synthesized dNTPαS and NTPαS analogs is excellent for direct DNA polymerization and RNA transcription,respectively.Since Klenow DNA polymerase and T7 RNA polymerase accept the Sp diastereomers of dNTPαS and NTPαS analogs,respectively,while the Rp diastereomers are neither substrates nor inhibitors,the diastereomerically-pure PS-DNAs and PS-RNAs can be conveniently synthesized enzymatically.

Phosphate-Methylated Oligonucleotides Past, Present and Future

At the moment, we see a great interest for application of Anti Sense Oligonucleotides (ASOs) in order to regulate the expression of genes related to certain diseases. These nucleotides obtained a number of fascinating properties by means of chemical manipulation of natural DNA and RNA under conservation of Watson-Crick base-pairing. About 35 years ago for our research in this field, we selected synthetically (short) phosphate-methylated DNA and RNA. It was concluded that there is an exclusive selection in hybridization affinity with natural DNA and RNA. These (bio)chemical and physical-chemical properties are extensively published. ASOs have found their way in public health as is clearly shown in the treatment of (progressive) neurological diseases. We focus specifically on the past, present and future of the phosphate-methylated oligonucleotides, illustrated with different research studies in chemistry and biophysics. A new field of application of modified DNAs is based on interactive improvements of sensitivity and specificity of nanowire field effect transistor gene chip by designing phosphate-methylated DNA as probe.

Modeling DNA ＆ RNA mutation using mset and topology

Mathematical equations are now found not only in the books, but also they help in finding solutions for the biological problems by explaining the technicality of the current biological models and providing predictions that can be validated and complemented to experimental and clinical studies. In this research paper, we use the mset theory to study DNA ＆： RNA mutations to discover the mutation occurrence. Also, we use the link between the concept of the meet and topology to determine the compatibility or similarity between ＂types＂, which may be the strings of bits, vectors, DNA or RNA sequences, etc.

Chemical and structural biology of nucleic acids and protein-nucleic acid complexes for novel drug discovery

Since nucleic acids(DNA and RNA) play very important roles in cells,they are molecular targets of many clinically used drugs,such as anticancer drugs and antibiotics.Because of clinical demands for treating various deadly cancers and drug-resistant strains of pathogens,there are urgent needs to develop novel therapeutic agents.Targeting nucleic acids hasn’t been the mainstream of drug discovery in the past,and the lack of 3D structural information for designing and developing drug specificity is one of the main reasons.Fortunately,many important structures of nucleic acids and their protein complexes have been determined over the past decade,which provide novel platforms for future drug design and discovery.In this review,we describe some useful nucleic acid structures,particularly their interactions with the ligands and therapeutic candidates or even drugs.We summarize important information for designing novel potent drugs and for targeting nucleic acids and protein-nucleic acid complexes to treat cancers and overcome the drug-resistant problems.

Synthesis and antiviral activity of some novel indole-2-carboxylate derivatives

A series of novel indole-2-carboxylate derivatives were synthesized and assayed to determine their in vitro broad-spectrum antiviral activities.The biological results showed that some of the synthesized compounds exhibited potent broad-spectrum antiviral activity.Notably,compound 8f showed the highest SI value(17.1)to Cox B3 virus.Compound 14f showed both potent inhibitory activity against influenza A(IC_(50)=7.53μmol/L)and the highest SI value(12.1).SAR results showed that the alkyloxy at the 4-position of indole ring was not crucial to the antiviral activities.Incorporation of an acetyl substituent at the amino group disfavored antiviral activity towards RNA viruses.

Separation axioms under crossover operator and its generalized

The purpose of this work is to construct a new crossover operator using the properties of DNA and RNA by using topological concepts in constructing flexible mathematical models in the field of biomathematics. Also, we investigate and study topological prop- erties of the constructed operators and the associated topological spaces of DNA and RNA. Finally we use the process of exchange for sequence of genotypes structures to construct new types of topological concepts to investigate and discuss several examples and some of their properties.