PCR-SSCP

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1 - DNA Polymorphisms at Candidate Gene Loci and Their Relation with Milk Production Traits in Murrah Buffalo (Bubalus bubalis)
دی.اس. کاله بی.آر. یاداو جی. پراساد

DNA polymorphism within diacylglycerol transferase 2 (DGAT2) / monoacyl glycerol transferases 2 (MOGAT2), leptin and butyrophilin genes were analysed using PCR-SSCP in Murrah buffalo. The single strand conformation polymorphism (SSCP) analysis of amplified gene fragment أکثر

DNA polymorphism within diacylglycerol transferase 2 (DGAT2) / monoacyl glycerol transferases 2 (MOGAT2), leptin and butyrophilin genes were analysed using PCR-SSCP in Murrah buffalo. The single strand conformation polymorphism (SSCP) analysis of amplified gene fragment in exon 5 of MOGAT2, exon 3 of leptin and intron 1 of butyrophilin gene revealed different patterns. A, B and C showed the following frequencies for each candidate gene in 53 (A=0.49, B=0.36 and C=0.15), 65 (A=0.38 and B=0.62) and 55 samples (A=0.6, B=0.31 and C=0.09) from Murrah buffaloes, respectively. The strand conformation polymorphism (SSCP) followed by DNA sequencing revealed one single nucleotide polymorphism (SNP) that was (c.193T>C) in MOGAT2, one single nucleotide polymorphism (SNP) (c.25 T>C) in leptin and one single nucleotide polymorphism (SNP) (c.184C>T>G) in butyrophilin gene confirmed BTI1 SSCP.The statistical analysis using general linear model procedure for association study, indicated that Murrah buffalo monoacyl glycerol transferases 2 (MOGAT2)(c.193T>C) single nucleotide polymorphism (SNP)and (c.25 T>C) in leptingenotypes were not significantly different (P>0.01) in Murrah buffalo for milk production traits milk yield, fat percentage and single nucleotide polymorphism (SNP)percentage. However, the statistical analysis for association study indicated BTI1 SSCP was significantly (P≤0.05) associated with 305-days lactation milk yield. The Murrah buffaloes with BTI1BB genotypes had 683.93 kg and 320.48 kg higher milk yield as compared to BTI1AA and BTI1CC genotypes, respectively. The positive association of butyrophilin single strand conformation polymorphism (SSCP) polymorphism with milk yield will be a useful tool for future selection and breeding strategies and genetic improvement of buffaloes for milk yield. تفاصيل المقالة

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2 - Identification of Complex Vertebral Malformation Carriers in Holstein and Guilan Native Cow Breeds in Iran Using SSCP Markers
H. Alaie S.Z. Mirhoseini M. Mehdizadeh S.B. Dalirsefat

Complex vertebral malformation(CVM) isanautosomal recessivehereditarydisorder caused bya point mutation in position 559 inexon 4 of the SLC35A3 gene on chromosome 3 inHolstein dairy cattle.This mutationchanges the function of uridine 5-diphosphate-N-acetylglucosamine tr أکثر

Complex vertebral malformation(CVM) isanautosomal recessivehereditarydisorder caused bya point mutation in position 559 inexon 4 of the SLC35A3 gene on chromosome 3 inHolstein dairy cattle.This mutationchanges the function of uridine 5-diphosphate-N-acetylglucosamine transporter protein bythe substitution of valine for phenylalanine at position 180 of this protein. The disease causes premature birth, aborted fetuses and stillborn calves. Latentrecessivegenesinheterozygousindividualscan be identified withhigh accuracy and repeatability using PCR-SSCP technique. Inthe present study,blood samples fromtwodifferent cow populations,including 100 Holstein cows and100 Guilan native cattle were randomly collected. Specific primers were used to amplify the 177-bp fragment of exon 4 of the SLC35A3 gene. No heterozygous genotype was detected in the studied samples. The Lack of carriers could be a consequence of the selection against the defective gene and preventative programs for entering mutant genes into the populations or very low frequency of this gene in these populations. However, there is a risk for increased genetic defects prevalence and it is necessary to develop screening programs to identify the defective gene. تفاصيل المقالة

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3 - Allelic Variation of MYF5 Gene Detected in the Camelus bactrianus
ن. هدایت-ایوریق س.ر. میرایی-آشتیانی م. مرادی شهر بابک و. واحدی ح. عبدی

The myogenic factors (MYF) 5 gene has been reported to contribute to muscle growth and development, therefore they are considered as candidate genes for growth and meat quality related traits. The MYF5 gene is expressed during proliferation of myoblasts and comprises 3 أکثر

The myogenic factors (MYF) 5 gene has been reported to contribute to muscle growth and development, therefore they are considered as candidate genes for growth and meat quality related traits. The MYF5 gene is expressed during proliferation of myoblasts and comprises 3 exons. To ascertain whether there is any variation in the camel MYF5 gene, we have used a polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) method for the analysis. In this study, coding region (exon 1) of the MYF5gene was investigated. Four unique SSCP patterns were detected in exon 1. Two Single nucleotide polymorphisms (SNPs) were detected - A/G and G/A in 98 and 366 position, respectively that create four haplotype, related to banding patterns. The variations detected in this study leads to a serin/asparagine and tryptophan/stop codon amino acid changes, respectively and could be considered for the development of gene-assisted selection in camel breeding. تفاصيل المقالة

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4 - CSN1S1 Gene: Allele Frequency, and the Relationship with Milk Production Traits in Three Indigenous Cattle Breeds and Holstein
S. Zakizadeh E.M. Prinzenberg M. Reissmann S.R. Miraei Ashtiani P. Reinecke G. Erhardt

CSN1S1is one of the major genes encoding milk proteins of mammals. In this study we determined allele frequencies of CSN1S1-5` flanking region as well as exon 17 variants and their effects on milk traits in three indigenous cattle breeds Mazandarani, Golpaygani (Bos ind أکثر

CSN1S1is one of the major genes encoding milk proteins of mammals. In this study we determined allele frequencies of CSN1S1-5` flanking region as well as exon 17 variants and their effects on milk traits in three indigenous cattle breeds Mazandarani, Golpaygani (Bos indicus) and Sarabi (Bos taurus) and Holstein cattle in Iran. CSN1S1*B variant was nearly fixed in Holstein but ranged from 0.40 to 0.66 in indigenous breeds. CSN1S1*C allele had higher frequency inindigenous breeds, especially inBos indicus. Four genetic variants of the promoter were found in all breeds in different frequencies with allele 2 being the prevalent in all breeds (frequency 0.359 to 0.711) and allele 4 the least frequent (0.074 to 0.011). Allele B of the coding region was found in combination with all four promoter alleles. Allele 4 of the promoter was not found in any cow having the exon 17 allele C in all breeds except Mazandarani. BC / 23 genotype yielded the highest fat percentage (P<0.05) in Holstein but it had no significant effect in Golpaygani. There was not any homozygous CSN1S1*CC cow, to investigate the influence of C variant for fat content. None of the genetic combinations had significant effect on fat yield, although variant '2' of promoter indicated a negative effect. No significant effect among various combined genotypes on milk yield was found, but CSN1S1*B tended to higher milk production. Differences of allelic frequencies and milk production traits found among these breeds might be due to differences in origin of breeds or selection breeding programs. تفاصيل المقالة

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5 - Study of DGAT1 Gene Polymorphisms with Carcass Traits in Iranian Zel and Lori-Bakhtiari Sheep Breeds
م. صادقی م. مخبر م. مرادی-شهر بابک و. سلطانی م. بهروزلک

Diacylglycerol acyltransferase1 (DGAT1) plays an important role in the metabolism of triglycerides which catalyze the ﬁnal step of triglyceride synthesis in animals. The objective of this study was to investigate the single-nucleotide polymorphisms (SNPs) in 5'UTR, exon أکثر

Diacylglycerol acyltransferase1 (DGAT1) plays an important role in the metabolism of triglycerides which catalyze the ﬁnal step of triglyceride synthesis in animals. The objective of this study was to investigate the single-nucleotide polymorphisms (SNPs) in 5'UTR, exon-1, and exon-2 of DGAT1 in two Iranian indigenous sheep breeds. A total of 309 animals including fat-tailed Lori-Bakhtiari (n=152) and thin-tailed Zel (n=157) were used in this study. The genotypic patterns were detected by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). Five SSCP patterns were detected for 5'UTR and exon-1 fragment by PCR-SSCP and subsequently confirmed by sequencing PCR products. The sequencing results revealed that there are three novel polymorphisms in 5'UTR and exon-1fragment of DGAT1 at the studied breeds. Out of the detected polymorphisms only A277G substitution in exon-1 of DGAT1 leads to the changes in amino acids (p.Arg26Gly). There was significant correlation (p < 0.05) between fat-tail weight (FTW) and back-fat thickness (BFT) and the observed genotypes in Lori-Bakhtiari breed; therefore, animals with G5 pattern had higher FTW and BFT compared to G1 pattern. The G1 and G5 genotypic patterns or haplotypes were different at their position 101 of 5'UTR region. No significant relationship (p < 0.05) was found between the detected genotypes of 5'UTR and exon-1 fragment of DGAT1 in Zel breed and carcass traits. These results revealed that detected DGAT1 novel SNPs had significant effects on carcass traits and they can be used as a marker for these traits. تفاصيل المقالة

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6 - Myostatin Gene Polymorphism and Its Association with Production Traits in Western Azerbaijan Native Chickens
S. Zandi P. Zamani K. Mardani

In the present study, the polymorphism of myostatin gene (MSTN) in native chickens of Western Azerbaijan Rearing and Breeding Institute was investigated. The blood samples were collected from eighty two randomly selected hens. Genomic DNA was extracted from blood sample أکثر

In the present study, the polymorphism of myostatin gene (MSTN) in native chickens of Western Azerbaijan Rearing and Breeding Institute was investigated. The blood samples were collected from eighty two randomly selected hens. Genomic DNA was extracted from blood samples and a fragment of myostatin including 599 bp in promoter and exon 1 was amplified using PCR method. Breeding values for body weight and carcass traits were predicted by univariate animal mixed model analysis, using WOMBAT software. The effects of different SSCP genotypes on breeding and phenotypic values of the studied traits were evaluated by general linear model analysis. Three different single strand conformational polymorphism(SSCP) genotypes as AA, AB and AC were identified, with frequencies of 0.244, 0.549 and 0.207, respectively. Shannon and Nei gene diversity indices and number of effective alleles in the studied population were 0.88, 0.53 and 2.2, respectively, which indicated a high diversity of the studied population. Moreover, the studied population was not in Hardy-Weinberg equilibrium. The effect of the SSCP genotypes on breeding and phenotypic values was significant only in the case of breeding value for body weight at 12 weeks of age, whereas, the AC genotype individuals, significantly (P<0.05) had the lowest breeding value for body weight at 12 weeks of age. Based on the results obtained, it could be concluded that the studied fragment of myostatin gene is polymorphic in native chickens of Azerbaijan and could be used for marker assisted selection. تفاصيل المقالة

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7 - Polymorphisms in GDF9 Gene and Its Relationship with Litter Size in Five Breeds of Black Goats
G.Q. Zhu Q.I. Wang Y.G. Kang Y.Z. Lv B.Y. Cao

In this study, we investigated the relationship between the genetic polymorphism of growth differentiation factor 9 (GDF9) genes and the litter size in 384 individuals of five breeds of black goats. Four pairs of primers were designed to detect single nucleotide polymor أکثر

In this study, we investigated the relationship between the genetic polymorphism of growth differentiation factor 9 (GDF9) genes and the litter size in 384 individuals of five breeds of black goats. Four pairs of primers were designed to detect single nucleotide polymorphism of GDF9 gene in goats by PCR-SSCP. The least square was used to analyze the relation between different genotypes and the litter size. The results showed that the PCR products from primer pair 1 (P1) displayed polymorphisms in three genotypes (AA, AB and BB) in big foot (BF) and Jintang (JT) black goats. For primer pairs of P2, P3 and P4, there was no polymorphism. The sequencing results revealed that there was a single nucleotide mutation (A792→G) in exon 2 of GDF9 gene in BF and JT black goats, and this mutation resulted in an amino acid change: valine→isoleucine. In BF and JT black goats, the average litter size in the third parity was significantly higher in genotype AA than both genotypes of AB and BB while the average little size of genotype AB was higher than that of genotype BB in the same parity. GDF9 gene could be therefore considered as a candidate gene for marker-assisted selection of litter size trait in goats. تفاصيل المقالة

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8 - Effect of Single Nucleotide Polymorphisms in IGF-1R Gene on Growth Rate Traits in Makooei Sheep
م. پسندیده ق. رحیمی و. همتی

Insulin-like growth factor 1 receptor (IGF-1R) is a main receptor of IGFs family which plays a critical role in the postnatal growth and skeletal growth in many species. However, there are few reports of IGF-1R gene structure and its effects on growth traits in sheep. T أکثر

Insulin-like growth factor 1 receptor (IGF-1R) is a main receptor of IGFs family which plays a critical role in the postnatal growth and skeletal growth in many species. However, there are few reports of IGF-1R gene structure and its effects on growth traits in sheep. The objectives of this study were detection of IGF-1R polymorphisms and assessment of their associations with growth traits in Iranian Makooei Sheep. Hence, 200 Makooei lambs were genotyped through polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP). The studied traits were birth weight (BW), weaning weight (WW), 6 months weight (6MW), average daily gains from birth to 3 months (ADG0-3), from 3 months to 6 months (ADG3-6), from birth to 6 months (ADG0-6) and corresponding Kleiber ratios (KR0-3, KR3-6, KR0-6).For this genetic position, three types of banding patterns (AA, AB and BB) were identified with the frequencies of 0.69, 0.16 and 0.15, respectively. In this study, IGF-1R genotypes indicated the significant associations with 6MW, ADG0-6, KR0-3 (P<0.05) and ADG0-3 (P<0.01). In all of the significant traits, The AAgenotype was linked to the highest values, while the BBgenotype was linked to the lowest values. The results of this study indicated that single nucleotide polymorphism (SNP) variation in IGF-1R gene can be used as a molecular marker for improving of growth traits in marker assisted selection programs in sheep. تفاصيل المقالة

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9 - بررسی ارتباط چهار پلی مورفیسم در ژن TNP2 با روند اسپرماتوژنز و ایجاد ناباروری مردان
الهام سیاسی احمد آل یاسین جواد مولا

در اسپرماتوژنز انسان، پروتامین ها به عنوان پروتئین های متصل شونده به DNA اسپرم جایگزین هیستون ها می شوند. برای تمایز و بلوغ اسپرم جایگزینی هیستون ها با پروتامین ها ضروری است. آسیب در ژن های کد کننده پروتامین ها می تواند منجر به تخریب اسپرماتوژنز و عدم تشکیل اسپرم کامل أکثر

در اسپرماتوژنز انسان، پروتامین ها به عنوان پروتئین های متصل شونده به DNA اسپرم جایگزین هیستون ها می شوند. برای تمایز و بلوغ اسپرم جایگزینی هیستون ها با پروتامین ها ضروری است. آسیب در ژن های کد کننده پروتامین ها می تواند منجر به تخریب اسپرماتوژنز و عدم تشکیل اسپرم کامل شده و در نتیجه سبب ناباروری مردان گردد. در این تحقیق به بررسی حضور چهار پلی مورفیسم در ژن TNP2 از ژن های خانواده پروتامین ها و ارتباط آن با ایجاد ناباروری ادیوپاتیک در مردان ازواسپرمی و الیگواسپرمی ایرانی پرداخته شده است. حضور چهار پلی مورفیسم T1019G،G1272C، G deletion at 1036 and 1046در ژن TNP2، پس از استخراج DNA از نمونه خون 96 مرد ازواسپرم و اولیگو اسپرم با ناباروری ادیوپاتیک و 100 نفر گروه کنترل با روش PCR-RFLP و PCR-SSCP بررسی شد و سپس با روش Sequencing نتایج تایید شد.فراوانی ژنوتایپ CC از پلی مورفیسم G1272C بین گروه بیمار و کنترل تفاوت نشان داد ولی با آنالیز آماری اختلاف معنی دار بین حضور این پلی مورفیسم در مقایسه گروه بیمار با کنترل مشاهده نشد (P>0/05). همچنین پلی مورفیسم های T1019G و G deletion at 1036 and 1046در هیچ یک از گروه بیمار و کنترل مشاهده نشد. مقایسه نتایج این تحقیق با مطالعات انجام شده در این زمینه نشان داد که بین حضور این چهار پلی مورفیسم بررسی شده در ایجاد ازواسپرمی و اولیگواسپرمی با ناباروری ادیوپاتیک در جمعیت مردان ایرانی ارتباط معنی داری وجود ندارد. تفاصيل المقالة

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