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Objective To screen and optimize the microsatellite DNA primers of the laboratory common marmoset, analyze and evaluate the population genetic quality for the marmosets (Callithrix jacchus) introduced into the Institute of Medical Laboratory Animal science, Chinese Academy of Medical Sciences. Methods A total of 30 marmosets were randomly chosen, and their genome DNA from blood was extracted using phenol/chloroform method. The microsatellite DNA was amplified using standard polymerase chain reaction (PCR). The amplification products were tested by STR scanning after 2% agrose gel and 8% PAGE electrophoresis. The data processing and genetic analysis were completed using the Popgene1. 32 software. Results A total of 20 pairs of microsatellite loci showed genetic polymorphism, and 147 alleles were detected. The number of allele was 5 to 10, average 7. 35. The effective allele was 2. 2500 to 6. 3830, average 4. 0402. The observed heterozygosity was 0. 000 to 0. 4667, average 0. 1533. The expected heterozygosity was 0. 1424 to 0. 4350, average 0. 2506. The Shannon diversity index was 1. 2242 to 2. 0324, average 1. 5949. The polymorphic information content was 0. 5366 to 0. 8254, average 0. 7053. Conclusions The 20 pairs of marmoset microsatellite primers are genetically highly diverse and are in a Hardy-Weinberg equilibrium.
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Objective To analyze the genetic quality of 24 domestic inbred strains mice using microsatellite loci panel.Methods Previously selected 30 microsatellite loci of mouse with high polymorphism and more allele numbers were used to synthesize corresponding fluorescently-labeled primers.Then the genomic DNA samples of each mouse were amplified by PCR and the products were analyzed by STR scanning to genotype the inbred strains of mice.Results Out of the 24 inbred strains, 15 inbred strains showed the same genotype within one strain at 30 loci.Among different strains, microsatellite loci indicated polymorphism which could be used to distinguish different strains.However, the rest 9 strains demonstrated polymorphism within strains.Conclusions Our stuoly provides a useful microsatellite panel to detect genetic quality of inbred mice and distinguish different strains with the optimized microsatellite loci.
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Objective Analysis of the genetic structure stability of Brandt ’ s vole ( lasiopodomys brandtii ) in closed colony using microsatellite marker technology.Methods Genomic DNA was extracted from tail tip using high-concentration-salt precipitation methods.Marked with fluorescent tags( Fam) , 7 microsatellite primers were filtered out by PCR, and the DNA structure of three consecutive generations of Brandt’ s vole was analyzed by microsatellite marker. Results By the analysis of the average heterozygosity and polymorphism information content, Brandt’ s vole populations maintaineda closed group of qualified genetic structure.Conclusions The present results show that the closed group of Brandt’ s vole species in our laboratory maintain a stable genetic structure.
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The genetic structure of small-scale landscape groups of Oncomelania hupensis in Songzi City ,Hubei Province was identify in this study .O .hupensis snails were collected from 10 habitats in Songzi City ,of which 6 polymorphic microsat-ellite DNA loci (T6-17 ,P101 ,D11 ,B14 ,T4-33 ,and C22) were carried out with GeneScan .The number of alleles (Na) ,het-erozygosity (H) ,fixation index (FST) of snails in each group ,genetic distance between groups ,and the polymorphic informa-tion content (PIC) were calculated .Cluster analysis was then carried out based on genetic distance ,and hierarchical AMOVA calculation was conducted .By certified the shells of snails ,10 groups were divided into light and ribbed shell (including shallow rib and deep rib) .There were 141 alleles in total detection on 10 populations and 20-34 alleles in each locus ,which were detec-ted for 23 .5 on average .The average number of alleles in 6 loci was 1 .575 and the number of alleles in each locus was uneven , showing large numerical differences ranged from 0 .445 to 3 .060 .The average observed heterozygosity (Ho) ranged from 0 .438 ue between paired populations was from -0 .015 64 to 0 .252 47 ,and the polymorphic information content in the population ranged from 0 .528 -0 .857 ,showing a high polymorphism .Hierarchical AMOVA calculations showed that inter-individual variation of the snails occupied 88 .4% of the total variations .Cluster analysis revealed that the three ribbed shell population in Munu Kou Village ,Hengti Village and Yixing Village first clustered to the three light shell population in Mashizizu Village , Mingzhu Village and Tuqiao Village ,then clustered to the light shell population in Tuanshan Village and Jiama Cao Village with the two shallow rib population in Desheng Village and Tianmu He Village .Under the different landscape environment of Songzi Area ,there were different shells presenting on the morphology of O .hupensis .Although there was a rich diversity on O .hupensis of Songzi City ,the genetic differences mostly present in individuals .Different groups didn’t show the significant genetic differentiation among the different shell morphology of O .hupensis .
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Objective To analyze and evaluate the population genetic quality of outbred KM mice from National Rodent Seed Center ( Shanghai ) .Methods A total of 30 outbred KM mice were randomly chosen.The genetic characteristics of the population were determined by PCR and STR scanning using 30 selected microsatellite loci. Popgen1.32 software was used to process the data.Results Thirty microsatellite loci shared 123 alleles in the KM mouse population.The average effective allele number and the average heterozygosity were 2.3989 and 0.5342, respectively.The average polymorphism information content ( PIC ) was 0.4735.Conclusions The outbred KM mouse population of Shanghai Seed Center has genetic stability and genetic diversity, and is satisfied with the genetic characteristics of closed colony laboratory animal.
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Objective To isolate the microsatellite DNA sequences of Oncomelania hupensis and analyze the polymorphic microsatellite loci.Methods The digested genomic fragments were hybridized with biotinylated oligonucleotide probes.The target fragments moleculars were captured and enriched.Then these fragments were cloned and sequenced.The suitable microsatellite loci were chosen and the polymorphism was screened by PAGE gel electrophoresis.Results A total of 205 microsateilite DNA sequences were obtained (GenBank accession numbers :GU204044～GU204248).The percentage of perfect microsatellite DNA sequence was 36.10% (74/205),with imperfect sequence as 49.76% (102/205) and compound sequence as 14.15% (29/205).Twenty typical microsatellite sequences were selected to design amplifying primers,and 13 microsatellite loci were found to be polymorphism.Conclusion A total of 205 microsatellite DNA sequences of Oncomelania hupensis are isolated and first reported,which will be useful for population genetic and mapping studies of Oncomelania hupensis.
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We determined the genetic diversity and distance between Jeju and Thoroughbred horses by genotyping for 20 microsatellite loci consisting of (TG)n repetitive sequence. The expected heterozygosity ranged from 0.1 to 0.789 in the Jeju horses and from 0.505 to 0.824 in the Thoroughbred horses. Polymorphic information content (PIC) values ranged from 0.09 to 0.709 in the Jeju horses and 0.365 to 0.730 in Thoroughbred horses. There were no significant differences in heterozygosity and PIC values between Jeju and Thoroughbred horsesHowever, LEX035 was estimated relatively high heterozygosity (0.789) and PIC value 0.709) in Jeju horses and LEX050 was respectively 0.824, and 0.730 in the Thoroughbred horses. We may conclude that the genetic differentiation was low between Jeju and Thoroughbred horses. LEX 050, LEX055, LEX059 and LEX 063 could be used as geneticmarkers for differentiating Jeju from Thoroughbred horses.
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DNA , Genetic Markers , Genetic Variation , Horses , Korea , Microsatellite Repeats , Polymorphism, Genetic , Repetitive Sequences, Nucleic AcidABSTRACT
In this study, microsatellite markers, developed for Alligator mississipiensis and Caiman latirostris, were used to assess parentage among individuals from the captive colony of Caiman latirostris at the University of São Paulo, in Piracicaba, São Paulo, Brazil. Many of the females in the colony were full siblings, which made maternal identification difficult due to genotypic similarity. Even so, the most likely mother could be identified unambiguously among offspring in most of the clutches studied. Two non-parental females displayed maternal behavior which would have misled managers in assigning maternity based on behavior alone. This set of variable loci demonstrates the utility of parentage testing in captive propagation programs.
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Background Familial hyperaldosteronism type Ⅱ(FH-Ⅱ) is characterized by autosomal dominant inheritance and hyperaldosteronism.Linkage analysis demonstrated chromosome 7q22 most likely harbor the genetic defect.Objective To identify the disease locus in a Chinese pedigree with FH-Ⅱ using genetic linkage analysis.Methods Haplotypes of the FH-Ⅱ pedigree were analyzed using four microsatellite markers(D7S531,D7S2521,D7S511,D7S481) at 7p22 to determine the critical region,and multipoint logarithm of odds(LOD) scores were calculated to assess linkage with FH-Ⅱ.Results Several members(affected members) in the family had hypertension,hypokalemia,hyperaldosteronism,low renin activity.Hypertension and hypokalemia was improved by spironolactone test.Dexamethasone suppression test was performed but without clinical or hyperaldosteronism improvement in three affected members,confirmed FH-Ⅱ family pedigree.The same haplotypes D7S531(ac,n=24)-D7S2521(ac,n=11)-D7S511(ca,n=21)-D7S481(ac,n=16) were shared by all known affected members and one suspicious member,but also by one unaffected member of the family,making it unlikely that this region contains the causative gene mutation.In this FH-Ⅱ family,the LOD scores of the four markers were
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Objective To isolate the microsatellite DNA of Phlebotomus chinensis and screen the polymorphic loci.Methods Genomic DNA fragments were hybridized with biotinylated oligonucleotide probes(AAT)17,(GA)25,(CCT)17,and(TG)18.The hybridized fragments were captured with Vectrex Avidin D and enriched by centrifugal ultrafiltration with ultra-4-column ultrafiltrate.The target fragments were amplified,cloned and sequenced.The suitable microsatellite loci were chosen in Ph.chinensis's library to establish PCR amplification assay.The polymorphism screening was conducted by PAGE with Ph.chinensis field specimens.Results The enrichment protocol used in this study was efficient,with a percentage of recombinant clone as 78.6%.There were 118 microsatellite sequences in library,the GenBank accession numbers were from FJ919812 to FJ919932(except GenBank accession numbers FJ919833,FJ919836,and FJ919869).There were 72 typical microsatellite sequences occupying 61.0% and the rest were 46 nontypical microsatellite sequences in the library.Twenty-two loci were chosen to polymorphism screening and PAGE showed that 14 loci were polymorphic.The loci of dinucleotide repeat were more polymorphic than those of trinucleotide and polynucleotide repeat.Conclusion The microsatellite-containing library of Ph.chinensis has been constructed with 118 sequences,and 14 new polymorphic microsatellite loci are reported.