This study was carried out the animal production department, genetic engineering lab, college of agriculture, （UoB）, Iraq. The aim of this study was to use the polymerase chain reaction restriction fragment length p...This study was carried out the animal production department, genetic engineering lab, college of agriculture, （UoB）, Iraq. The aim of this study was to use the polymerase chain reaction restriction fragment length polymorphism （PCR-RFLP） as a fast, efficient and low cost method to detect the genetic variants of kappa-casein gene （k-CN） in Iraqi buffalo using three different primers specific for bovine k-CN to amplify the gene segment, followed by digestion using restriction enzyme （Hind III） for genotyping. DNA from 50 Iraqi buffaloes was extracted by phenol chloroform method. PCR was carried out in a final reaction volume of 25 μL and the reaction mixture was subjected to standard PCR protocol. The results of this work show that among the examined 50 Iraqi Buffalo were homozygous for the K-CN and genotyped as BB for all three primers but gave different bands. Thus PCR-RFLP using Hind III revealed all the samples to be monomorphic for this locus. The restriction digestion analysis of 397 bp PCR product of k-CN indicates the presence of two fragments of 154 bp and 225 bp for BB-genotype. A 437 bp fragment of the bovine genomic K-CN gene was amplified. One Hind III restriction site is found in position 346 of the amplified fragment of allele k-CN B, yielded 91 bp and 346 bp. Amplified products from Iraqi buffalo （530）, after being digested with Hind III, yielded two separate DNA fragments of different sizes i.e., 160 bp and 370 bp. For the first time completed research such specifications in Iraq, for the first time using molecular biology in genetic identification. Our objectives of this study have been to aid in understanding domestication, Buffalo origin and their history and evolution, to identify genetically unique breeds, to provide an objective basis for conservation decisions and to aid the formulation of breeding plans.展开更多
Intraocular pressure elevation can induce retinal ganglion cell death and is a clinically reversible risk factor for glaucoma,the leading cause of irreversible blindness.We previously demonstrated that casein kinase-2...Intraocular pressure elevation can induce retinal ganglion cell death and is a clinically reversible risk factor for glaucoma,the leading cause of irreversible blindness.We previously demonstrated that casein kinase-2 inhibition can promote retinal ganglion cell survival and axonal regeneration in rats after optic nerve injury.To investigate the underlying mechanism,in the current study we increased the intraocular pressure of adult rats to 75 mmHg for 2 hours and then administered a casein kinase-2 inhibitor(4,5,6,7-tetrabromo-2-azabenzimidazole or 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole)by intravitreal injection.We found that intravitreal injection of 4,5,6,7-tetrabromo-2-azabenzimidazole or 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole promoted retinal ganglion cell survival and reduced the number of infiltrating macrophages.Transcriptomic analysis showed that the mitogen activated protein kinase signaling pathway was involved in the response to intraocular pressure elevation but was not modulated by the casein kinase-2 inhibitors.Furthermore,casein kinase-2 inhibition downregulated the expression of genes(Cck,Htrsa,Nef1,Htrlb,Prph,Chat,Slc18a3,Slc5a7,Scn1b,Crybb2,Tsga10ip,and Vstm21)involved in intraocular pressure elevation.Our data indicate that inhibition of casein kinase-2 can enhance retinal ganglion cell survival in rats after acute intraocular pressure elevation via macrophage inactivation.展开更多
Tiller number and grain size are important agronomic traits that determine grain yield in rice.Here,we demonstrate that DEFECTIVE TILLER GROWTH 1(DTG1),a member of the casein kinase 1 protein family,exerts a co-regula...Tiller number and grain size are important agronomic traits that determine grain yield in rice.Here,we demonstrate that DEFECTIVE TILLER GROWTH 1(DTG1),a member of the casein kinase 1 protein family,exerts a co-regulatory effect on tiller number and grain size.We identified a single amino acid substitution in DTG1(I357K)that caused a decrease in tiller number and an increase in grain size in NIL-dtg1.Genetic analyses revealed that DTG1 plays a pivotal role in regulation of tillering and grain size.The DTG1^(I357K) allelic variant exhibited robust functionality in suppressing tillering.We show that DTG1 is preferentially expressed in tiller buds and young panicles,and negatively regulates grain size by restricting cell proliferation in spikelet hulls.We further confirm that DTG1 functioned in grain size regulation by directly interacting with Grain Width 2(GW2),a critical grain size regulator in rice.The CRISPR/Cas9-mediated elimination of DTG1 significantly enhanced tiller number and grain size,thereby increasing rice grain yield under field conditions,thus highlighting potential value of DTG1 in rice breeding.展开更多
文摘This study was carried out the animal production department, genetic engineering lab, college of agriculture, （UoB）, Iraq. The aim of this study was to use the polymerase chain reaction restriction fragment length polymorphism （PCR-RFLP） as a fast, efficient and low cost method to detect the genetic variants of kappa-casein gene （k-CN） in Iraqi buffalo using three different primers specific for bovine k-CN to amplify the gene segment, followed by digestion using restriction enzyme （Hind III） for genotyping. DNA from 50 Iraqi buffaloes was extracted by phenol chloroform method. PCR was carried out in a final reaction volume of 25 μL and the reaction mixture was subjected to standard PCR protocol. The results of this work show that among the examined 50 Iraqi Buffalo were homozygous for the K-CN and genotyped as BB for all three primers but gave different bands. Thus PCR-RFLP using Hind III revealed all the samples to be monomorphic for this locus. The restriction digestion analysis of 397 bp PCR product of k-CN indicates the presence of two fragments of 154 bp and 225 bp for BB-genotype. A 437 bp fragment of the bovine genomic K-CN gene was amplified. One Hind III restriction site is found in position 346 of the amplified fragment of allele k-CN B, yielded 91 bp and 346 bp. Amplified products from Iraqi buffalo （530）, after being digested with Hind III, yielded two separate DNA fragments of different sizes i.e., 160 bp and 370 bp. For the first time completed research such specifications in Iraq, for the first time using molecular biology in genetic identification. Our objectives of this study have been to aid in understanding domestication, Buffalo origin and their history and evolution, to identify genetically unique breeds, to provide an objective basis for conservation decisions and to aid the formulation of breeding plans.
基金supported by the National Natural Science Foundation of China,Nos.81570849,81100931the Natural Science Foundation of Guangdong Province of China,Nos.2015A030313446,2020A1515011413(all to LPC).
文摘Intraocular pressure elevation can induce retinal ganglion cell death and is a clinically reversible risk factor for glaucoma,the leading cause of irreversible blindness.We previously demonstrated that casein kinase-2 inhibition can promote retinal ganglion cell survival and axonal regeneration in rats after optic nerve injury.To investigate the underlying mechanism,in the current study we increased the intraocular pressure of adult rats to 75 mmHg for 2 hours and then administered a casein kinase-2 inhibitor(4,5,6,7-tetrabromo-2-azabenzimidazole or 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole)by intravitreal injection.We found that intravitreal injection of 4,5,6,7-tetrabromo-2-azabenzimidazole or 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole promoted retinal ganglion cell survival and reduced the number of infiltrating macrophages.Transcriptomic analysis showed that the mitogen activated protein kinase signaling pathway was involved in the response to intraocular pressure elevation but was not modulated by the casein kinase-2 inhibitors.Furthermore,casein kinase-2 inhibition downregulated the expression of genes(Cck,Htrsa,Nef1,Htrlb,Prph,Chat,Slc18a3,Slc5a7,Scn1b,Crybb2,Tsga10ip,and Vstm21)involved in intraocular pressure elevation.Our data indicate that inhibition of casein kinase-2 can enhance retinal ganglion cell survival in rats after acute intraocular pressure elevation via macrophage inactivation.
基金supported by the Sichuan Science and Technology Support Project (2022ZDZX0012,2021YFYZ0016,2023YFN0007,2021YFYZ0027)the National Natural Science Foundation of China (32171966,U23A20180).
文摘Tiller number and grain size are important agronomic traits that determine grain yield in rice.Here,we demonstrate that DEFECTIVE TILLER GROWTH 1(DTG1),a member of the casein kinase 1 protein family,exerts a co-regulatory effect on tiller number and grain size.We identified a single amino acid substitution in DTG1(I357K)that caused a decrease in tiller number and an increase in grain size in NIL-dtg1.Genetic analyses revealed that DTG1 plays a pivotal role in regulation of tillering and grain size.The DTG1^(I357K) allelic variant exhibited robust functionality in suppressing tillering.We show that DTG1 is preferentially expressed in tiller buds and young panicles,and negatively regulates grain size by restricting cell proliferation in spikelet hulls.We further confirm that DTG1 functioned in grain size regulation by directly interacting with Grain Width 2(GW2),a critical grain size regulator in rice.The CRISPR/Cas9-mediated elimination of DTG1 significantly enhanced tiller number and grain size,thereby increasing rice grain yield under field conditions,thus highlighting potential value of DTG1 in rice breeding.
