CRISPR/Cpf1 has emerged recently as an effective tool for genome editing in many organisms,but its use in pigs to generate precise genetic modifications has seldom been described.Myostatin(MSTN)is a well-characterized...CRISPR/Cpf1 has emerged recently as an effective tool for genome editing in many organisms,but its use in pigs to generate precise genetic modifications has seldom been described.Myostatin(MSTN)is a well-characterized negative regulator of muscle development,and natural mutations in this gene cause a double-muscled phenotype in many species.However,to the best of our knowledge,no naturally occurring mutation in MSTN has been found in pigs.In addition,no living pig models with sophisticated modifications orthologous to natural mutations in MSTN have yet been reported.In this study,we exploited the CRISPR/Cpf1 system to introduce a predefined modification orthologous to the natural MSTN mutation found in Belgian Blue cattle(thus known as the Belgian Blue mutation).Our research demonstrated that the cutting efficiency of CRISPR/Cpf1 was 12.3%in mixed porcine fetal fibroblasts in drug free medium,and 41.7%in clonal colonies obtained using G418 selection.Then,the Cpf1-sgRNA vector,ssODN template,and a self-excision cassette were co-transfected into porcine fetal fibroblasts.After G418 selection,8 clonal colonies were examined and 5 with genetic modification were found.Of these 5,2 harbored the precise 11-bp deletion.Using 1 heterozygous clonal colony,2 cloned Duroc piglets were successfully generated,which was heterozygous for the Belgian Blue mutation.In summary,our results demonstrate that CRISPR/Cpf1 system can be used efficiently to generate double-stranded breaks,and also to mediate homologous recombination to introduce precise genomic modifications in pigs.展开更多
Increasing consumer awareness of health and climate is being reflected in their dietary choices.This is reflected in the strong growth in the market for plant-based food,with plant-based burgers(PBB)being the most pop...Increasing consumer awareness of health and climate is being reflected in their dietary choices.This is reflected in the strong growth in the market for plant-based food,with plant-based burgers(PBB)being the most popular meat alternative.PBB must be based on natural ingredients that increase nutritional value,improve technological properties,and create satisfying physical sensations.Fibers obtained from agri-food by-products can potentially meet all of these requirements.The dietary fiber concentrate(DFC)produced from the forced roots of Belgian endive(Cichorium intybus var.foliosum)(by-product)has shown potential for application in PBB.Replacing soy protein with 5%and 10%of DFC from Belgian endive(DFC-BE)resulted in lower caloric value and sugar content and doubled dietary fiber concentration in the end product,enough to justify a“high fiber”nutrition claim(>6 g per 100 g).Baking properties such as baking yield,moisture retention,and dimeter reduction also improved.Application of DFC-BE can also be used to regulate pH,color,texture profile,and sensory attributes.Individual recipes must be adjusted to obtain the required results.展开更多
Dogs are used for centuries as people helpers. One of possible use is a sport and service. BSM （Belgian Shepherd Malinois） and GS （German shepherd） dogs are most used breeds like service dogs and also most represe...Dogs are used for centuries as people helpers. One of possible use is a sport and service. BSM （Belgian Shepherd Malinois） and GS （German shepherd） dogs are most used breeds like service dogs and also most represented at sports competitions. Our aim was to compare the results of these breeds in the World Cup of International tests of working dogs （in years 2003-2011）, which is the peak of competitions. The authors evaluated results of German shepherd dog performance test and Belgian shepherd Malinois performance in disciplines such as tracking, obedience and defense. Belgian Shepherds achieved in all disciplines higher score than German shepherds. It was found out that Belgian shepherd Malinois get significant higher score in all disciplines （tracking, obedience and defense） than German shepherd. However, both of breeds are used successfully as service dogs.展开更多
为了研究大型肉牛比利时蓝牛生长发育的遗传规律,筛选优异基因,试验基于Illumina Bovine SNP 50K芯片数据,采用PLINK软件对270头比利时蓝牛常染色体数据进行基因组长纯合片段(ROH)检测并基于选择信号分析,通过核苷酸多态性检测取前5%的...为了研究大型肉牛比利时蓝牛生长发育的遗传规律,筛选优异基因,试验基于Illumina Bovine SNP 50K芯片数据,采用PLINK软件对270头比利时蓝牛常染色体数据进行基因组长纯合片段(ROH)检测并基于选择信号分析,通过核苷酸多态性检测取前5%的单核苷酸多态性(single nucleotide polymorphism,SNP)位点,基于牛参考基因组(ARS-UCD1.2)对结果SNPs进行基因注释,对候选基因进行GO功能注释与KEGG信号通路富集分析,并计算染色体上ROH长度占基因组总长度的比例(FROH)。结果表明:在全部270个个体数据中共检测出1893个ROH片段,平均长度13.2311 Mb,平均FROH为0.0392;得到与生长发育相性状相关的基因有NEB、TET2、NEK11、NCKAP1、MYH15、EIF4A2、bta-miR-1248-1、DCAF8、PRORP、DOCK3、SYT15、MYEF2、ZDHHC13,与公牛生育能力相关的基因有CFAP61、DNAL1、BAG1。说明通过对比利时蓝牛生长发育性状相关分子标记的解析可以为比利时蓝牛遗传改良提供理论指导。展开更多
为了解榆林市散养比利时兔球虫的感染情况,选取榆林市某散养比利时兔场采用饱和盐水漂浮法和重铬酸钾培养法对采集的散养比利时兔粪便样品进行感染情况检查。结果表明,散养比利时兔球虫平均感染率为62.3%,其中成年兔感染率是57.6%,感染...为了解榆林市散养比利时兔球虫的感染情况,选取榆林市某散养比利时兔场采用饱和盐水漂浮法和重铬酸钾培养法对采集的散养比利时兔粪便样品进行感染情况检查。结果表明,散养比利时兔球虫平均感染率为62.3%,其中成年兔感染率是57.6%,感染强度为500;幼龄兔感染率是71.4%,感染强度为800。经检测发现8种球虫,分别为盲肠艾美尔球虫(E.coecicola Cheissin,1947)、纳格浦尔艾美尔球虫(E.nagpurensis Guil et Ray,1966)、斯氏艾美尔球虫(E.stiedae Kiss kalt et Hartmann,1907)、梨形艾美尔球虫(E.piriformis Kotlan et Pospesch,1934)、肠艾美尔球虫(E.intestinalis Cheissin,1948)、马氏艾美尔球虫(E.matsubayashii Tsunoda,1952)、小型艾美尔球虫(E.imeriaexigua Yakimoff,1934)、中型艾美尔球虫(E.mebia Kcsscl,1929),其中盲肠艾美尔球虫、纳格浦尔艾美尔球虫、斯氏艾美尔球虫为优势虫种,所占比例分别为25.3%、23.1%、14.3%。本调查结果可为榆林市防治散养比利时兔球虫病提供一定的数据参考和理论依据。展开更多
为了解兔肉质性状特征,给兔的品种选种选育提供理论依据,选择UCP1基因设计1对引物扩增其第3外显子及第2内含子部分序列,采用DNA池PCR反应,直接测序法快速检测比利时兔、新西兰兔和加利福尼亚兔3个兔品种UCP基因多态性。结果在新西兰兔...为了解兔肉质性状特征,给兔的品种选种选育提供理论依据,选择UCP1基因设计1对引物扩增其第3外显子及第2内含子部分序列,采用DNA池PCR反应,直接测序法快速检测比利时兔、新西兰兔和加利福尼亚兔3个兔品种UCP基因多态性。结果在新西兰兔中发现1个多态性位点(T387C),多态位点对UCP1基因的RNA结构产生影响,其最小自由能由-917.51 k J/mol变为-927.12 k J/mol,而其他2个品种没有检测到该突变。展开更多
基金supported by the National Transgenic Breeding Program of China (2016ZX08006001)the Doctor’s Fund of Southwest University, China (SWU 118082)
文摘CRISPR/Cpf1 has emerged recently as an effective tool for genome editing in many organisms,but its use in pigs to generate precise genetic modifications has seldom been described.Myostatin(MSTN)is a well-characterized negative regulator of muscle development,and natural mutations in this gene cause a double-muscled phenotype in many species.However,to the best of our knowledge,no naturally occurring mutation in MSTN has been found in pigs.In addition,no living pig models with sophisticated modifications orthologous to natural mutations in MSTN have yet been reported.In this study,we exploited the CRISPR/Cpf1 system to introduce a predefined modification orthologous to the natural MSTN mutation found in Belgian Blue cattle(thus known as the Belgian Blue mutation).Our research demonstrated that the cutting efficiency of CRISPR/Cpf1 was 12.3%in mixed porcine fetal fibroblasts in drug free medium,and 41.7%in clonal colonies obtained using G418 selection.Then,the Cpf1-sgRNA vector,ssODN template,and a self-excision cassette were co-transfected into porcine fetal fibroblasts.After G418 selection,8 clonal colonies were examined and 5 with genetic modification were found.Of these 5,2 harbored the precise 11-bp deletion.Using 1 heterozygous clonal colony,2 cloned Duroc piglets were successfully generated,which was heterozygous for the Belgian Blue mutation.In summary,our results demonstrate that CRISPR/Cpf1 system can be used efficiently to generate double-stranded breaks,and also to mediate homologous recombination to introduce precise genomic modifications in pigs.
基金the CichOpt project which is funded in the frame of the ERA-NET FACCE SURPLUS,FACCE SURPLUS has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No 652615 and VLAIO,the Flemish agency for innovation and entrepreneurship.
文摘Increasing consumer awareness of health and climate is being reflected in their dietary choices.This is reflected in the strong growth in the market for plant-based food,with plant-based burgers(PBB)being the most popular meat alternative.PBB must be based on natural ingredients that increase nutritional value,improve technological properties,and create satisfying physical sensations.Fibers obtained from agri-food by-products can potentially meet all of these requirements.The dietary fiber concentrate(DFC)produced from the forced roots of Belgian endive(Cichorium intybus var.foliosum)(by-product)has shown potential for application in PBB.Replacing soy protein with 5%and 10%of DFC from Belgian endive(DFC-BE)resulted in lower caloric value and sugar content and doubled dietary fiber concentration in the end product,enough to justify a“high fiber”nutrition claim(>6 g per 100 g).Baking properties such as baking yield,moisture retention,and dimeter reduction also improved.Application of DFC-BE can also be used to regulate pH,color,texture profile,and sensory attributes.Individual recipes must be adjusted to obtain the required results.
文摘Dogs are used for centuries as people helpers. One of possible use is a sport and service. BSM （Belgian Shepherd Malinois） and GS （German shepherd） dogs are most used breeds like service dogs and also most represented at sports competitions. Our aim was to compare the results of these breeds in the World Cup of International tests of working dogs （in years 2003-2011）, which is the peak of competitions. The authors evaluated results of German shepherd dog performance test and Belgian shepherd Malinois performance in disciplines such as tracking, obedience and defense. Belgian Shepherds achieved in all disciplines higher score than German shepherds. It was found out that Belgian shepherd Malinois get significant higher score in all disciplines （tracking, obedience and defense） than German shepherd. However, both of breeds are used successfully as service dogs.
文摘为了解榆林市散养比利时兔球虫的感染情况,选取榆林市某散养比利时兔场采用饱和盐水漂浮法和重铬酸钾培养法对采集的散养比利时兔粪便样品进行感染情况检查。结果表明,散养比利时兔球虫平均感染率为62.3%,其中成年兔感染率是57.6%,感染强度为500;幼龄兔感染率是71.4%,感染强度为800。经检测发现8种球虫,分别为盲肠艾美尔球虫(E.coecicola Cheissin,1947)、纳格浦尔艾美尔球虫(E.nagpurensis Guil et Ray,1966)、斯氏艾美尔球虫(E.stiedae Kiss kalt et Hartmann,1907)、梨形艾美尔球虫(E.piriformis Kotlan et Pospesch,1934)、肠艾美尔球虫(E.intestinalis Cheissin,1948)、马氏艾美尔球虫(E.matsubayashii Tsunoda,1952)、小型艾美尔球虫(E.imeriaexigua Yakimoff,1934)、中型艾美尔球虫(E.mebia Kcsscl,1929),其中盲肠艾美尔球虫、纳格浦尔艾美尔球虫、斯氏艾美尔球虫为优势虫种,所占比例分别为25.3%、23.1%、14.3%。本调查结果可为榆林市防治散养比利时兔球虫病提供一定的数据参考和理论依据。
文摘为了解兔肉质性状特征,给兔的品种选种选育提供理论依据,选择UCP1基因设计1对引物扩增其第3外显子及第2内含子部分序列,采用DNA池PCR反应,直接测序法快速检测比利时兔、新西兰兔和加利福尼亚兔3个兔品种UCP基因多态性。结果在新西兰兔中发现1个多态性位点(T387C),多态位点对UCP1基因的RNA结构产生影响,其最小自由能由-917.51 k J/mol变为-927.12 k J/mol,而其他2个品种没有检测到该突变。
