1.A Speedy Extraction Method for Bacterial Chromosomal DNA
Wei ZHENG ; Chun-Shan QUAN ; Yong-Zhe PIAO ; Sheng-Di FAN ;
China Biotechnology 2006;0(04):-
How to get functional gene from uncultured-microbiology is the hotspot content of microbial ecology. What the most important is how to obtain the pure and integrated genomic DNA. An efficient, nonselective extraction method to gain chromosomal DNA from eight kinds of bacteria was introduced. Amount DNA released by hot-detergent gave the highest DNA yields from different G + and G- bacteria. Running 20 hours by PFGE mode, the size of total DNA is over 23kb. The pure DNA could be digested by Hind Ⅲ and used in PCR. The total environmental DNA also can be extracted from soil by the same method. As a result it showed a new way for the environmental DNA extraction.
2.The Extraction Method of Bacterial DNA from Activated Sludges
Wei ZHENG ; Chun-Shan QUAN ; Yong-Zhe PIAO ; Jun-Hua WANG ; Sheng-Di FAN ;
China Biotechnology 2006;0(07):-
Methods for studying the population diversity of microorganism in activated sludge usually require enrichment of bacterial genome.The efficient information on microbial species composition provided and shifted in diversity revealed are dependent on the effective DNA recovery technique.The method was based on washing by alkaline phosphate buffer and digestion with extended heating of the activated sludge suspension in the presence of lysozyme and freeze-thawing in high-salt-SDS buffer.The extraction was tested for four activated sludge differing in places and dates.The DNA fragment from all sludge was integrity.DNA yields ranged from 105 to 823 ?g/g sludge and were of sufficient purity for PCR-based 16S ribosomal DNA analysis and restriction digested.In general,all methods produced DNA pure were not enough for PCR amplification and libraries construction.As basis of experimental goals,the study provides an appropriate extraction method of microbial DNA in sludge.
3.Genetic polymorphism of the PRODH and 5-HT(2A) receptor gene in Korean-Chinese and Han nationality in Chinese Yanbian area.
Wan-kui JIANG ; Qing-song XU ; Yong-quan PIAO
Chinese Journal of Medical Genetics 2008;25(1):93-95
OBJECTIVETo investigate the polymorphisms distribution of genes related with mental diseases, such as proline dehydrogenase gene (PRODH) and serotonin 2A(5-HT(2A)) receptor gene, among Korean-Chinese and Han nationality in Chinese Yanbian area.
METHODSBy utilizing techniques of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), the polymorphisms for -1945 region of the PRODH gene and -1438 region of the 5-HT(2A) receptor gene were analyzed.
RESULTSThe PRODH genotype frequencies of MM, Mm and mm in Korean-Chinese population were 61.4%,34.4%,4.2% respectively, the allele frequencies were 0.786 for M and 0.214 for m respectively, and those in Han population were 55.2%,38.5%,6.3%, 0.745 and 0.255 respectively. The 5-HT(2A) receptor gene genotype frequencies of AA, AG and GG in Korean-Chinese population were 15.6%,67.7%,16.7% respectively, the allele frequencies were 0.495 for A and 0.505 for G respectively, and those in Han population were 11.5%,65.6%,22.9%, 0.443 for A and 0.557 for G.
CONCLUSIONAll of the genotype distribution of both loci in Chinese Korean and Han nationality in Yanbian area meet Hardy-Weinberg equilibrium, and show stronger ability in human identity. The data obtained can be used in human identity, paternity testing and Chinese Korean ethnic group study.
Adult ; Animals ; China ; Electrophoresis ; Ethnic Groups ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Korea ; ethnology ; Male ; Mental Disorders ; genetics ; Mice ; Polymorphism, Genetic ; Proline Oxidase ; genetics ; Receptor, Serotonin, 5-HT2A ; genetics