1.Analysis of endophytic fungi community of Ligusticum chuanxiong using PCR-DGGE.
Hai WANG ; Zhu-Yun YAN ; Dong-Mei HE ; Meng WANG
China Journal of Chinese Materia Medica 2013;38(12):1893-1897
OBJECTIVETo research the diversity of endophytic fungal communities among Ligusticum chuanxiong growing at 5 areas in Sichuan province, and illuminate the developing mechanism of geoherbs from the microecological perspective.
METHODThe PCR-DGGE and DNA sequencing techniques were used to analyze the endophytic fungi community of L. chuanxiong.
RESULTThe community of endophytic fungi present difference among different growing areas. Though minor difference were found among individuals at the same area, similarity among individuals from the same growing areas were higher significantly than those from different growing areas. Compared with the other 4 growing areas, L. chuanxiong from Shiyang town, Dujiangyan city had more abundant endophytic fungi and low similarity to others, and which probably had special types of fungi.
CONCLUSIONThe abundant and stable endophytic fungal community is an important factor for the development of geoherb L. chuanxiong at Shiyang town, Dujiangyan city.
Denaturing Gradient Gel Electrophoresis ; methods ; Ligusticum ; genetics ; growth & development ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA
2.A preliminary analysis of changes in composition of intestinal microbiota during infancy using polymerase chain reaction-denaturing gradient gel electrophoresis.
Shan-Shan LI ; Jun-Jie MIAO ; Zi-Xi WU ; Jian-Rong YAO ; Ming LI ; Qian YU ; Fang HE
Chinese Journal of Contemporary Pediatrics 2017;19(3):331-336
OBJECTIVETo investigate the composition of bacteria in the stools of infants and the colonization of intestinal microbiota during infancy.
METHODSFresh stools were collected from 15 healthy infants at 0, 2, 4, 7, 10, 14, and 28 days and 3, 6, and 12 months after birth. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the composition of intestinal microbiota, perform sequencing of dominant bacteria, and to analyze the changes in the composition of intestinal microbiota during infancy.
RESULTSDGGE fingerprint showed that the composition of intestinal microbiota during infancy changed significantly over time after birth. The cloning and sequencing results indicated that Proteobacteria colonized the earliest, mainly the obligate aerobes Enterobacter and Pseudomonas, followed by the obligate anaerobes (Clostridium hathewayi and Veillonella parvula) and the facultative anaerobe Clostridium ramosum in Firmicutes, and Verrucomicrobia. Actinobacteria colonized the latest, mainly Bifidobacterium, and gradually became dominant bacteria.
CONCLUSIONSDuring infancy, obligate aerobes colonize the intestinal tract the earliest, followed by obligate anaerobes and facultative anaerobes. Proteobacteria colonizes the earliest, followed by Firmicutes and Verrucomicrobia, and Actinobacteria, mainly Bifidobacterium, colonizes the latest.
Denaturing Gradient Gel Electrophoresis ; methods ; Feces ; microbiology ; Female ; Gastrointestinal Microbiome ; Humans ; Infant ; Male ; Polymerase Chain Reaction ; methods
3.Polymorphism study of small nuclear ribonucleoprotein polypeptide N gene rs220030 by DGGE.
Yun ZHAO ; Hong-Mei XU ; Zi-Qin ZHAO
Journal of Forensic Medicine 2011;27(3):186-188
OBJECTIVE:
To analyze the polymorphism of rs220030, a SNP which is located in the promoter region of small nuclear ribonucleoprotein polypeptide N (SNRPN) gene in the Chinese Han population and to obtain the data of population genetics.
METHODS:
The denaturing gradient gel electrophoresis (DGGE) method was applied to detect the polymorphism of rs220030 in 100 unrelated and healthy individuals from the Shanghai Han population. The genotyping result of this SNP was confirmed by TaqMan assay in some typical samples.
RESULTS:
DGGE results showed 4 bands for CT heterozygote, and 1 band for CC or TT homozygote, and those results were confirmed by The TaqMan SNP genotyping assays. Genotyping results showed 34 individuals with CC, 41 with CT and 25 with TT of rs220030. The allele frequencies for C and T were 0.545 and 0.455, respectively. H was 0.500, PIC was 0.373, DP was 0.654, and PE was 0.186. The distribution of genotype frequencies were in Hardy-Weinberg equilibrium.
CONCLUSION
DGGE is a quick and effective method in the analysis of SNP polymorphism in small population. Statistical parameters of rs220030 for forensic evaluation meet the requirements for forensic identification and paternity testing.
Alleles
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Asian People/genetics*
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China/ethnology*
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DNA Primers
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Denaturing Gradient Gel Electrophoresis/methods*
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Gene Frequency
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Genetic Markers
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Genetics, Population
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Genotype
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Heterozygote
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Humans
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Polymerase Chain Reaction
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Polymorphism, Single Nucleotide/genetics*
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Promoter Regions, Genetic
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snRNP Core Proteins/genetics*