Application of Molecular Biology Technology in Identification of Modern Mongolian Medicine

The application of molecular biology technology in the identification and quality control of Mongolian medicine is increasing gradually,and it provides a new method for identifying fake and inferior products and confused products of Mongolian medicine.In this paper,the application and prospect of molecular biology technology(such as DNA barcoding and PCR molecular identification technique)in the identification of crude Mongolian medicine were reviewed.

Advances in Application of PCR Amplification in Etiologic Diagnosis

In recent years,with the rapid development of molecular biology diagnostic technology,many new polymerase chain reaction(PCR)technologies with high specificity and good sensitivity have gradually been developed.While expanding the range of detection methods,these technologies inevitably have some disadvantages.Therefore,in clinical pathogen diagnosis,medical personnel should choose the detection method according to the detection purpose and pathogen characteristics.In this paper,the basic principle,application scope,advantages and disadvantages and development of various emerging PCR diagnostic techniques are respectively described in order to provide a theoretical reference for the selection of pathogenic biological diagnostic techniques in the clinical practice.

Cloning of TRP Gene of Bombyx mori

[Objective]The aim was to clone Bombyx mori TRP gene.[Method]The total RNA of Bombyx mori was extracted during its pupal period with Trizol method,and then the TRP gene was cloned by RT-PCR.[Result]The full-length cDNA of TRP gene was successfully obtained,and the ORF fragment（858 bp） of TRP gene was cloned by PCR.[Conclusion]TRP gene of Bombyx mori was cloned for the first time,which could provide solid basis for the study on its function.

Influence of 6-BA on Incidence Rate of Chinese Cabbage Clubroot (Plasmodiophora brassicae Woron)

[Objective] The paper was to study the relationship between cytokinin and Chinese cabbage clubroot, and to explore the incidence mechanism of clubroot. [Method]The spores were extracted from the tumors of diseased plant, and their DNA was extracted for PCR test. The diagnosed spores were inoculated to the culture soils in erlenmeyer flask at seeding stage, at seed germination and 21 d after sowing, and 0.08 μmol/L 6-BA was added to the soil at germination. Spores were extracted from the root tumors developed in inoculated plants and examined by scanning electronic microscope （ SEM）, and the incidence rate of clubroot in 6-BA treatment and control was recorded. [ Result] Plasmodiophora brassicae Woron could cause the clubroot incidence of Chinese cabbage planted in erlenmeyer flasks, the incidence rate of chibroot in the treatment adding with 0.08 μmol/L 6-BA was 100%, while that in the treatment without 6-BA was 57%, and the volume of former tumor was much larger than the latter. SEM showed that the size of resting spore of P. brassicae was 1.5 -4.3μm. [ Conduslon] 6-BA applied at germination could significantly promote the formation of tumor of Chinese cabbage clubroot.

The Study of Food-Grade Induced Expression and Enzymatic Properties of L-Arabinose Isomerase from Lactobacillus plantarum WU14 with High D-Tagatose Yield

L-arabinose isomerase (L-AI) is the key enzyme for D-galactose isomerization of D-tagatose by biological method. In this research, Lactobacillus plantarum WU14 with high D-tagatose yield was identified as Lactobacillus plantarum was isolated from the number of lactic acid bacteria from pickled vegetables. The crude L-arabinose isomerase activity of Lactobacillus plantarum WU14 with high D-tagatose yield was 13.95 U/mL under the optimal temperature 60&degC, pH 7.17 and substrate concentration 0.8 mol/L, and the conversion rate of 56.12% could be gained after 28 hours. Protein structure and specific of L-Arabinose Isomerase of Lactobacillus plantarum WU14 were researched. The results showed that L-arabinose isomerase is mainly composed of alpha helix and random coil. Then the recombinant L-AI gene was inserted into the food-grade expression vector pRNA48 and expressed in L. lactis NZ9000 successfully. The target protein expression reached the maximum amount when the induced concentration of nisin reaches 30 ng/mL after 12 h. And the crude enzyme activity of recombinant bacteria reached 6.21 U/mL under 60&degC. Otherwise the optimal conversion rate recombinant of L. lactis NZ9000/pRNA48-L-AI can reach 39.21% under the temperature of 50&degC, pH 7.17 and D-galactose concentration was 0.6 mol/L.

RAPID DETECTION OF ONCOGENE AMPLIFICATION IN PARAFFIN SECTIONS OF BREAST CARCINOMAS USING QUANTITATIVE DIFFERENTIAL PCR AND FLUORESCENT DNA TECHNOLOGY

Gene amplification is a common mechanism of oncogene activation and contributes to tumor progression. Analysis of such genetic alterations are relevant to the understanding of tumor genetics and could provide prognostic information for the individual patient. Standard analytical approaches using Southern blot and slot blot require a large amount of good

Identification of Arbuscular mycorrhizal multiplicity in the saline-sodic soils

This study focused on the Arbuscular mycorrhizal(AM)fungal diversity in the saline-sodic soils based on native spore density and most probable number(MPN)assay.Identification through spore morphology showed existence of five genera in the various crop rhizospheres.The physico-chemical analysis of the native soils revealed that they were saline-sodic with pH ranging from(8.7±0.5)to(9.5±0.6)and habituated five different genera of AM fungi including Glomus,Scutellospora,Acaulospora,Sclerocystis and Gigaspora.Each location revealed presence of varied species of AM fungus namely Acaulospora and Glomus in rhizosphere of maize;Scutellospora and Glomus in tulsi;four isolates of Glomus in onion;Glomus and Sclerocystis in guava;three isolates of Glomus in rice;Glomus in neem and Gigaspora and Glomus in bamboo.The molecular identification through nested PCR analysis showed amplification of 600 bp size in SSU rDNA gene in samples A and C(predominated by Acaulospora and Glomus mosseae respectively).