1.Molecular characterization of cotton leaf Curl Multan virus and its satellite DNA that infects Hibiscus rosa-sinensis.
Ming-Jie MAO ; Zi-Fu HE ; Hao YU ; Hua-Ping LI
Chinese Journal of Virology 2008;24(1):64-68
Virus isolate G6 was obtained from Hibiscus rosa-sinensis showing yellow and leaf curl symptoms in Guangzhou, Guangdong Province. The complete nucleotide sequence of DNA-A was determined to be 2 737 nucleotides encoding six potential ORFs. Comparison showed that G6 DNA-A had more than 89% sequence identify with all isolates of Cotton leaf curl Multan virus (CLCuMV) and shared the highest sequence identify (96.1%) with CLCuMV isolate 62. G6 DNA-A had 87.1%-89.8% sequence identity with those of CLCuRV isolates, while less than 87% identities with other begomoviruses. Phylogenetic analysis of G6 DNA-A and selected begomoviruses showed that G6 was most closely related to CLCuMV isolates, and they clustered together as a separate branch. Satellite DNA molecule (G6 DNAbeta) was found to be associated with G6 using the primers beta01 and beta02. G6 DNAbeta contains 1346 nucleotides, with a potential functional ORF (C1) in complementary sense DNA. Pairwise comparison indicated that G6 DNAbeta had the highest sequence identities with CLCuMV DNAbeta (92.1%) and CLCuRV DNAbeta (88.7%), but less than 80% sequence identities with other reported satellite DNA molecules. Phylogenetic analysis indicated that G6 DNAbeta was most closely related to CLCuMV DNAbeta and the two DNAbetas clustered together as a separate branch, and formed the main branch with DNAbeta of CLCuRV and MYVV-Y47. It is concluded that G6 infecting Hibiscus rosa-sinensis is an isolate of CLCuMV.
Base Sequence
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DNA, Satellite
;
chemistry
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DNA, Viral
;
chemistry
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Geminiviridae
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classification
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genetics
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Gossypium
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virology
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Hibiscus
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virology
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Phylogeny
2.A validation study of four single locus probes (MS1, MS31, MS43 and g3) in a Korean population: further evaluation for paternity testing.
Journal of Korean Medical Science 1995;10(4):243-249
Hypervariable tandem repetitive regions in human DNA are proving to be increasingly useful for genetic analysis in humans. We chose four single locus probes (SLP; MS1, MS43, MS8 and g3) for a validation test among Koreans. The specimens were from 216 unrelated individuals and 33 paternity inclusion families. Extracted DNA from EDTA blood was restricted by Hinfl and electrophoresed in 0.7% agarose gel, transferred and hybridized with chemiluminescent probes. Heterozygosity was over 90% by all of the probes. Total numbers of unassignable mutant bands from 33 paternity inclusion cases were 5, and the highest mutation rate was determined in probe MS1(0.045). The probability of having the same DNA band between two unrelated individuals was 5.7 x 10(-10) when four SLPs were used at the same time. The data presented here on allele frequencies and mutation rates provide preliminary data supporting the validity of these probes in paternity analysis and forensic investigators in the Korean population.
Alleles
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Chromosome Mapping
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DNA, Satellite/*genetics
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Female
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Heterozygote
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Human
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Male
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Mutation
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Myoglobin/*genetics
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*Paternity
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Support, Non-U.S. Gov't
3.Role of multicolor fluorescence in situ hybridization (FISH) in simultaneous detection of probe sets for chromosome 18, X and Y in uncultured amniotic fluid cells.
Journal of Korean Medical Science 1999;14(4):438-442
Major aneuploidies diagnosed prenatally involve the autosomes 13, 18, and 21, and sex chromosomes. Fluorescence in situ hybridization (FISH) allows rapid analysis of chromosome copy number in interphase cells. The purpose of this study was to evaluate the role of multicolor fluorescence in situ hybridization in simultaneous detection of probe sets for chromosome 18, X, and Y in uncultured amniotic fluid cells as a safer alternative method for aneuploidy detection prenatally. Fifty amniotic fluid samples were analyzed by FISH and standard cytogenetics. Mean time to obtain results was three days for fluorescence in situ hybridization and 20 days for karyotype. Fluorescence in situ hybridization was informative in 43 samples (86%), and within this group, two aneuploidies were correctly identified. This evaluation demonstrates that FISH with X, Y, and 18 alpha satellite DNA probes could accurately and rapidly detect aneuploidies involving these chromosomes and could be used in any prenatal clinical laboratory.
Amniocentesis/methods*
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Amniotic Fluid/cytology
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Aneuploidy
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Centromere/genetics
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Chromosomes, Human, Pair 18*
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Color
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DNA Probes
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DNA, Satellite/analysis
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Female
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Human
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In Situ Hybridization, Fluorescence/methods*
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Karyotyping
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Pregnancy
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Sex Chromosome Abnormalities/genetics
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Sex Chromosome Abnormalities/diagnosis*
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X Chromosome*
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Y Chromosome*
4.Hypermethylation of hMLH1 and microsatellite instability in ovarian mucinous tumors.
Huai-zeng CHEN ; Da-feng YE ; Xing XIE ; Min HU ; Wei-guo LU
Acta Academiae Medicinae Sinicae 2003;25(4):457-461
OBJECTIVESTo investigate the role of hMLH1 promoter hypermethylation and microsatellite instability (MSI) in the development of ovarian mucinous tumors.
METHODSOne hundred and seven of paraffin-embedded specimens of ovarian mucinous tumors (malignant 49, borderline 35, and benign 23) were collected from Women's Hospital, School of Medicine, Zhejiang University from 1995 to 2001. The assessment of MSI was based on the use of a panel of six microsatellite markers (BAT-25, BAT-26, BAT-40, D5S346, D17S250, and D2S123) by polymerase chain reaction (PCR). Hypermethylation of hMLH1 promoter region was detected using restriction cut analysis.
RESULTS4.3% (1/23), 14.3% (5/35), and 36.7% (18/49) of benign tumors, borderline tumors, and malignant tumors respectively displayed hypermethylation of the hMLH1 promoter. The hMLH1 promoter hypermethylation rate of malignant group was significantly higher than that of borderline and benign group (P = 0.023, 0.004), but no significant difference between the borderline group and the benign group (P = 0.438); 4.3% (1/23), 8.6% (3/35), and 16.3% (8.49) of benign tumors, borderline tumors, and malignant tumors showed MSI positive phenotype. But there were no significant differences each other in the MSI positive phenotype rate; 75% (9/12) MSI positive phenotype ovarian mucinous tumors were hypermethylated at hMLH1 promoter, while the MSI-phenotype tumors were unmethylated in 84.2% (80.95) of cases. There was significant correlation between MSI positive phenotype and hMLH1 promoter hypermethylation (P = 0.000).
CONCLUSIONSIn ovarian mucinous tumors, malignant, borderline, and benign tumors exist hMLH1 promoter hypermethylation. Hypermethylation of hMLH1 promoter results MSI in ovarian mucinous tumors. Methylation of hMLH1 promoter and MSI may be involved in the carcinogenesis of ovarian mucinous cancer.
Adaptor Proteins, Signal Transducing ; Base Pair Mismatch ; Carrier Proteins ; Chromosomal Instability ; Cystadenocarcinoma, Mucinous ; genetics ; DNA Methylation ; DNA Repair ; DNA, Neoplasm ; genetics ; DNA, Satellite ; Female ; Genes, Neoplasm ; Humans ; Microsatellite Repeats ; genetics ; MutL Protein Homolog 1 ; Neoplasm Proteins ; genetics ; Nuclear Proteins ; Ovarian Neoplasms ; genetics ; Promoter Regions, Genetic ; genetics
5.Anticancer and antimicrobial activity of mangrove derived fungi Hypocrea lixii VB1.
B Valentin BHIMBA ; D A AGNEL DEFORA FRANCO ; Jibi Merin MATHEW ; Geena Mary JOSE ; Elsa Lycias JOEL ; M THANGARAJ
Chinese Journal of Natural Medicines (English Ed.) 2012;10(1):77-80
AIM:
Mangrove is one of the oldest living tree species and its leaves are among the most extensively studied botanicals in use today. Scientific research throughout the world has found evidence to support the fact that its foliar extracts have great potential against human microbial pathogens. This study highlights the isolation of foliar fungi from Rhizophora mucronata, Avicenna officialis and Avicenna marina.
METHOD:
It was isolated in Sabouroud's Dextrose Agar and mass cultivation was done in Sabouroud's Dextrose broth.
RESULTS:
The ethyl acetate extract showed maximum antibacterial activity which inturn checked for different concentration against bacterial pathogens and anticancer activity for Hep2 and MCF7 cell line in vitro. The DNA was isolated from the fungi and the ITS region of 5.8 s RNA was sequenced and assigned to new species as they are separated from the type strains phylogenetic neighbors by sequence similarities.
CONCLUSION
This preliminary screening of fungal endophytes revealed their potential to yield potent bioactive compounds for drug discovery programmes.
Anti-Bacterial Agents
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pharmacology
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therapeutic use
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Antineoplastic Agents
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pharmacology
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therapeutic use
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Avicennia
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microbiology
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Base Sequence
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Biological Products
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pharmacology
;
therapeutic use
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Cell Line, Tumor
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DNA, Fungal
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Endophytes
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Humans
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Hypocrea
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genetics
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MCF-7 Cells
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Neoplasms
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drug therapy
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Phylogeny
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Phytotherapy
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RNA, Satellite
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Rhizophoraceae
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microbiology
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Species Specificity