Somatic cells were prepared from sea snail enzyme digests of Porphyra yezoensis thalli. Us ing SDS - Proteinase K as extraction solution, total DNA was isolated from the somatic cells. The crude extracts of total DNA ...Somatic cells were prepared from sea snail enzyme digests of Porphyra yezoensis thalli. Us ing SDS - Proteinase K as extraction solution, total DNA was isolated from the somatic cells. The crude extracts of total DNA were purified with glassmilk, and the resulting DNA was of sufficient quality for digestion of restriction endonuclease. DNA bands were clearly observed in the restriction patterns of EcoRI, PstI and HaeIII respectively. The presence of DNA hands in the restriction pattern of total DNA indicated that the genome of Porphyra yezoensis may be small. Unexpectedly, using guanidinium isoth iocyanate and sarcosyl as extraction solution, a plasmid-like DNA band (2.3 Kb) was directly found in the isolated total DNA of Porphyra yezoensis. A very simple and convenient method for plasmid-like DNA isolation has been established.展开更多
[ Objective] The aim was to develop a fast and effective DNA extraction method of intestinal microflora, a modified method of chloroform extraction, and to provide the basis for quantitative and qualitative detection....[ Objective] The aim was to develop a fast and effective DNA extraction method of intestinal microflora, a modified method of chloroform extraction, and to provide the basis for quantitative and qualitative detection. [ Method] Through the improvement of conventional DNA extraction method, a rapid and efficient DNA extraction method was developed. Compared with the real-time PCR result of control sample and the result of QIAamp DNA Stool Mini kit, the developed method was verified. [ Result] The DNA yield of the developed method was 100 times as much as that of QIAamp DNA Stool Mini kit. And the real-time PCR result showed that the efficiency of DNA extraction of the developed method was higher than that of the QIAamp DNA Stool Mini kit. [ Conclusion] This modified method is inexpensive, efficient and rapid, and it is suitable for large quantities of feces samples.展开更多
Glycyrrhizic acid(GA) in Glycyrrhiza uralensis(G.uralensis) is physiologically active.In this study,the total DNA of wild G.uralensis was randomly transformed into Hansenula anomala by implantation of low-energy Ar^+ ...Glycyrrhizic acid(GA) in Glycyrrhiza uralensis(G.uralensis) is physiologically active.In this study,the total DNA of wild G.uralensis was randomly transformed into Hansenula anomala by implantation of low-energy Ar^+ and N^+,to produce five recombinant yeast strains relating to biological synthesis of the GA or Glycyrrhetinic acid (GAs).After culturing in liquid medium for 96 h,the resultant GA,18α-GAs and 18β-Gas were determined by reversed-phase high performance liquid chromatography(RP-HPLC),and the corresponding concentrations were 114.49,0.56,and 0.81 mg·L^(-1).After one hundred primers were analyzed with random amplified polymorphic DNA (RAPD),the seven different DNA fragments were produced by the N7059 strain of recombined yeasts,and,the polymerase chain reaction(PCR) verified that one of them came from the genome of G.uralensis,indicating a successful transfer of genetic information by ion implantation.展开更多
Total DNA is isolated from Porphyra cells which have been prepared from sea snail enzyme digests of Porphyra thalli. The crude extracts of total DNA are rapidly purified with Wizard DNA Clean-up resin by using modifie...Total DNA is isolated from Porphyra cells which have been prepared from sea snail enzyme digests of Porphyra thalli. The crude extracts of total DNA are rapidly purified with Wizard DNA Clean-up resin by using modified washing solution. DNA bands are clearly observed in the restriction endonuclease patterns of P. yezoensis and P. haitanensis total DNAs. Both P. yezoensis and P. haitanensis have specific DNA bands in the restriction patterns of EcoRI, PastI and HaeIII. According to the band pattern of total DNA, the discrimination between P. yezoensis and P. haitanensis is very easy especially in the restriction pattern of HaeIII. The comparative results indicate that the band pattern of total DNA is a new type and reliable DNA marker for identification and classification of Porphyra species.展开更多
文摘Somatic cells were prepared from sea snail enzyme digests of Porphyra yezoensis thalli. Us ing SDS - Proteinase K as extraction solution, total DNA was isolated from the somatic cells. The crude extracts of total DNA were purified with glassmilk, and the resulting DNA was of sufficient quality for digestion of restriction endonuclease. DNA bands were clearly observed in the restriction patterns of EcoRI, PstI and HaeIII respectively. The presence of DNA hands in the restriction pattern of total DNA indicated that the genome of Porphyra yezoensis may be small. Unexpectedly, using guanidinium isoth iocyanate and sarcosyl as extraction solution, a plasmid-like DNA band (2.3 Kb) was directly found in the isolated total DNA of Porphyra yezoensis. A very simple and convenient method for plasmid-like DNA isolation has been established.
文摘[ Objective] The aim was to develop a fast and effective DNA extraction method of intestinal microflora, a modified method of chloroform extraction, and to provide the basis for quantitative and qualitative detection. [ Method] Through the improvement of conventional DNA extraction method, a rapid and efficient DNA extraction method was developed. Compared with the real-time PCR result of control sample and the result of QIAamp DNA Stool Mini kit, the developed method was verified. [ Result] The DNA yield of the developed method was 100 times as much as that of QIAamp DNA Stool Mini kit. And the real-time PCR result showed that the efficiency of DNA extraction of the developed method was higher than that of the QIAamp DNA Stool Mini kit. [ Conclusion] This modified method is inexpensive, efficient and rapid, and it is suitable for large quantities of feces samples.
基金Supported by National Natural Science Foundation of China(Grant No.30560182,30960006)
文摘Glycyrrhizic acid(GA) in Glycyrrhiza uralensis(G.uralensis) is physiologically active.In this study,the total DNA of wild G.uralensis was randomly transformed into Hansenula anomala by implantation of low-energy Ar^+ and N^+,to produce five recombinant yeast strains relating to biological synthesis of the GA or Glycyrrhetinic acid (GAs).After culturing in liquid medium for 96 h,the resultant GA,18α-GAs and 18β-Gas were determined by reversed-phase high performance liquid chromatography(RP-HPLC),and the corresponding concentrations were 114.49,0.56,and 0.81 mg·L^(-1).After one hundred primers were analyzed with random amplified polymorphic DNA (RAPD),the seven different DNA fragments were produced by the N7059 strain of recombined yeasts,and,the polymerase chain reaction(PCR) verified that one of them came from the genome of G.uralensis,indicating a successful transfer of genetic information by ion implantation.
文摘Total DNA is isolated from Porphyra cells which have been prepared from sea snail enzyme digests of Porphyra thalli. The crude extracts of total DNA are rapidly purified with Wizard DNA Clean-up resin by using modified washing solution. DNA bands are clearly observed in the restriction endonuclease patterns of P. yezoensis and P. haitanensis total DNAs. Both P. yezoensis and P. haitanensis have specific DNA bands in the restriction patterns of EcoRI, PastI and HaeIII. According to the band pattern of total DNA, the discrimination between P. yezoensis and P. haitanensis is very easy especially in the restriction pattern of HaeIII. The comparative results indicate that the band pattern of total DNA is a new type and reliable DNA marker for identification and classification of Porphyra species.
