Curcumin, a major constituent of turmeric, corrects cystic fibrosis defects

Cystic fibrosis is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR). The most common mutation, DeltaF508, results in the production of a misfolded CFTR protein that is retained in the endoplasmic reticulum and targeted for degradation. Curcumin is a nontoxic Ca-adenosine triphosphatase pump inhibitor that can be administered to humans safely. Oral administration of curcumin to homozygous DeltaF508 CFTR mice in doses comparable, on a weight-per-weight basis, to those well tolerated by humans corrected these animals' characteristic nasal potential difference defect. These effects were not observed in mice homozygous for a complete knockout of the CFTR gene. Curcumin also induced the functional appearance of DeltaF508 CFTR protein in the plasma membranes of transfected baby hamster kidney cells. Thus, curcumin treatment may be able to correct defects associated with the homozygous expression of DeltaF508 CFTR.     

猪CFTR基因特异性锌指核酸酶的筛选与鉴定

【目的】获得靶向猪囊性纤维化跨膜传导调节因子(Cystic fibrosis transmembrane conductance regulator,CFTR)基因的特异锌指核酸酶(Zinc finger nuclease,ZFN),为建立CFTR基因敲除猪细胞系提供技术支持。【方法】通过开源式(Oligomerized pool engineering,OPEN)方法筛选,首先以已知三锌指蛋白为框架,随机突变单个锌指关键位点氨基酸编码序列,建立人工三锌指蛋白随机库;然后应用细菌双杂交技术,从库中筛选出能够结合CFTR基因靶位点的三锌指蛋白;最后将获得的锌指蛋白与非限制性核酸内切酶FokⅠ组装成特异ZFN,通过酵母验证体系,检测ZFN靶向切割其识别序列的效率。【结果】获得3个人工三锌指蛋白随机库,每个库约含有2×10⁶个单克隆,通过2轮细菌双杂交筛选,分别获得48个针对CFTR基因左右两侧靶位点的三锌指蛋白。ZFN活性验证结果表明,约90%的ZFN能够在酵母细胞内实现靶向切割,获得了高效特异的ZFN。【结论】获得了猪CFTR基因高效特异的ZFN。     

A mutation in PRKAG3 associated with excess glycogen content in pig skeletal muscle

A high proportion of purebred Hampshire pigs carries the dominant RN- mutation, which causes high glycogen content in skeletal muscle. The mutation has beneficial effects on meat content but detrimental effects on processing yield. Here, it is shown that the mutation is a nonconservative substitution (R200Q) in the PRKAG3 gene, which encodes a muscle-specific isoform of the regulatory gamma subunit of adenosine monophosphate-activated protein kinase (AMPK). Loss-of-function mutations in the homologous gene in yeast (SNF4) cause defects in glucose metabolism, including glycogen storage. Further analysis of the PRKAG3 signaling pathway may provide insights into muscle physiology as well as the pathogenesis of noninsulin-dependent diabetes mellitus in humans, a metabolic disorder associated with impaired glycogen synthesis.     

广西巴马小型猪JAK2基因外显子14克隆及SNP位点分析

【目的】克隆广西巴马小型猪非受体型酪氨酸蛋白激酶（JAK2）基因外显子14并进行SNP位点分析,为探讨广西巴马小型猪JAK2基因多态性与有关临床疾病治疗奠定基础。【方法】参照GenBank已发表的猪JAK2基因编码序列设计引物,克隆出猪JAK2基因外显子14,经测序鉴定后,利用PCR-SSCP技术对其进行SNP位点分析。【结果】PCR扩增获得的巴马小型猪JAK2基因外显子14序列为205 bp,与猪的同源性为100.0%,与人类的同源性为94.2%;猪和人类在JAK2基因外显子14序列中有6个位点不同;PCR-SSCP分析结果表明,JAK2基因外显子14不存在多态性。【结论】广西巴马小型猪JAK2基因外显子14不存在多态性,其基因纯合度高,是进行育种研究和医学试验的理想实验动物模型。     

基因编辑猪的研究现状

猪是重要的农业经济动物,猪肉是人类获得蛋白质营养物质的主要途径之一。与此同时,猪在解剖学、生理学及遗传背景、疾病特征等方面与人类极为相似。因此,猪既是农业动物生产性状改良的重要对象,又是人类疾病、异种器官移植等生物医学领域的研究对象。随着基因编辑技术的飞速发展,出现了越来越多操作简单、运用广泛且安全的新型编辑技术,可以快速获得单碱基编辑、基因敲除或敲入的细胞系,并通过体细胞克隆等技术获得基因编辑猪。综述了基因编辑猪的制备及其在农业及医学领域中的研究进展,以期为猪的农业生产性状改良和医学研究提供参考。     

Genetics of the behavior of domestic pigs toward humans

Hereditary determination and the reality of major gene control of inheritance of behavior having selection value in domestic pigs toward humans are shown.     

Cystic fibrosis locus defined by a genetically linked polymorphic DNA marker

A polymorphic DNA marker has been found genetically linked, in a set of 39 human families, to an autosomal recessive gene that causes cystic fibrosis (CF), a disease affecting one in 2000 Caucasian children. The DNA marker (called D0CRI-917) is also linked to the PON locus, which by independent evidence is linked to the CF locus. The best estimates of the genetic distances are 5 centimorgans between the DNA marker and PON and 15 centimorgans between the DNA marker and the CF locus, meaning that the location of the disease gene has been narrowed to about 1 percent of the human genome (about 30 million base pairs). Although the data are consistent with the interpretation that a single locus causes cystic fibrosis, the possibility of genetic heterogeneity remains. The discovery of a linked DNA polymorphism is the first step in molecular analysis of the CF gene and its causative role in the disease.     

猪CTSZ基因的克隆及真核表达载体的构建

组织蛋白酶Z基因(Cathepsin Z gene, CTSZ)是影响猪生长、肉质等性状的一个重要候选基因.本研究以长白猪的背最长肌为试验材料,采用RT-PCR的方法克隆CTSZ基因CDS序列全长,对该序列进行了生物信息学分析,并构建pEGFP-N1-CTSZ融合表达载体,通过脂质体介导瞬时转染猪成纤维细胞,30 h后进行荧光表达检测.结果本试验克隆出猪CTSZ基因CDS全序列912 bp,并成功构建pEGFP-N1-CTSZ融合表达载体,瞬时转染后在猪成纤维细胞中表现为绿色荧光蛋白表达.本试验为进一步研究该基因的生物学功能奠定了基础.     

高纤维日粮添加复合酶制剂对猪消化率与生产性能的影响

通过消化试验和生长试验,对猪饲料中常用的复合酶制剂的效果进行了研究。结果表明,酶制剂改善了15kg体重仔猪和60kg体重阶段生长猪的能量、粗蛋白和粗纤维的消化率,但只有体重15kg仔猪的粗纤维消化率有显著改善(p<0.05)。在生长猪阶段(30～60kg)添加酶制剂,猪的饲料转换效率极显著优于对照组(p<0.01),但对日增重无显著影响。而在肥育猪阶段(60～90kg),饲料中添加酶制剂对日增重和饲料转化率无显著影响。饲料酶制剂的应用效果与猪的生长阶段和酶制剂的组成密切相关。     

Gene engineering in swine for agriculture

Domestic pigs are the second most important source of meat world-wide, and the genetic improvement of economic traits, such as meat production, growth, and disease resistance, is a critical point for efficient production in pigs. Through conventional breeding and selection programs in pigs, which are painstakingly slow processes, some economic traits, such as growth and backfat, have been greatly improved over the past several decades. However, the improvement of many polygenetic traits is still very slow and challenging to be improved by conventional breeding strategies. The development of reproductive knowledge and a variety of techniques, including foreign gene transfer strategies, somatic cell nuclear transfer(SCNT) and particularly, recently developed nuclease-mediated genome editing tools, has provided efficient ways to produce genetically modified(GM) pigs for the dramatic improvement of economic traits. In this review, we briefly discuss the progress of genomic markers used in pig breeding program, trace the history of genetic engineering, mainly focusing on the progress of recently developed genome editing tools, and summarize the GM pigs which have been generated to aim at the agricultural purposes. We also discuss the specific challenges facing application of gene engineering in pig breeding, and future prospects.     

Production of homeobox A10 gene transgenic pigs by somatic cell nuclear transfer

Homeobox A10(Hoxa10) gene is one of the most important candidate genes associated with the reproductive performance of humans and mice. Overexpression of Hoxa10 in mouse endometrium can increase litter size. Moreover, Hoxa10 plays a key role in regulating the embryo implantation of sows. This study aimed to generate transgenic pigs using Hoxa10 via somatic cell nuclear transfer(SCNT). We established seven Hoxa10-transgenic cell lines, and two of the cell lines were selected as nuclear donors for the transfer. A total of 1 270 cloned embryos were generated and transferred to five surrogate mothers(Landrace×Yorkshire). Eight cloned male piglets were produced including one with cryptorchidism. Six transgenic piglets grew up healthy and produced 56 offspring. Finally, we obtained six transgenic male pigs and 26 transgenic positive offspring that can be used to further study the regulatory mechanism of Hoxa10 on the reproductive performance of pigs.     

historical review of pig testing programs in Denmark

Until1887pig breeding in Denmark wasbased upon the export to Germany of heavy pigs,weighing from120to150kg.To satisfy thismarket the old native Danish Landrace breed wascrossed with Middle White and Berkshire boarsimported from England in order to produce stillfatter pigs.In1887the importation of Danishpigs became prohibited by Germany and ourfarmers had to change over to the production ofbacon for the English market.This change over was not an easy one.Forthe production of bacon,a comple…     

论中国乡村休闲农业发展中的猪文化挖掘

养猪在中国祖先生活中占有重要的地位。在远古时期,祖先对猪产生图腾崇拜,在民间流传着许多关于猪的美好传说,也传承着多种多样的猪美食。分析中国乡村传统文化中的猪文化,并提出在休闲农业发展中传承猪文化的几点建议。     

转基因和基因编辑猪的研究进展

猪是重要的农业经济动物,其在遗传背景、解剖学、生理病理学、营养代谢和疾病特征等方面与人类高度相似,是农业生产性状改良、人类疾病动物模型和异种器官移植研究的重要对象。随着ZFN、TALEN和CRISPR等基因编辑技术的出现,转基因或基因编辑猪的研究取得快速发展。本文将结合当前我国转基因及基因编辑猪的研究现状,对基因编辑技术在农业生产性状改良如提高产肉量、改善肉质、提高抗病能力和建立人类疾病模型、动物生物反应器、异种器官移植等方面进行综述,以期为猪遗传改良和医学研究提供参考。     

猪Krox20基因的cDNA克隆与表达分析

根据已报道的人和鼠的Krox20基因,以同源性高的区域设计1对简并引物,以猪脂肪组织RNA为模板,通过RT-PCR方法,获得了猪的Krox20基因的部分cDNA,长度是938bp。多种组织的半定量RT-PCR研究表明,仔猪时期,该基因在心脏、肝脏、脾脏、大肠、小肠、脂肪组织和肌肉中均有表达,9月龄成年猪也在上述组织中有所表达,但是表达量不如仔猪,并且还在性腺中检测到该基因的表达。     

转基因猪研究新进展

由于猪既是重要的经济动物,又是常用的实验动物,并且因为其解剖、组织、生理和营养代谢等方面与人类相似,因此转基因猪的研究意义重大。纵观转基因猪的发展历史,关键核心技术包括显微注射技术、体细胞核移植技术、精子载体技术、胚胎干细胞介导技术、病毒转染技术、基因打靶、ZFN技术和TALENS技术。科学家们通过这些技术对猪进行了大量研究,已经在生命科学领域取得了令人欣慰的成就。     

吉林省水稻生产的碳足迹与水足迹时空变化特征

农业生产对全球气候变暖、水资源短缺和环境污染具有重要影响。碳足迹和水足迹分别是评估温室气体排放和水资源消耗的指标。采用生命周期评价方法,对吉林省水稻生产的碳足迹和水足迹进行核算,分析碳足迹和水足迹的时空变化特征及其构成。结果表明:2007—2017年吉林省水稻生产的碳足迹年均值为0.74 kg·kg^-1。甲烷排放是水稻生产碳足迹的主要组分,占比为41.55%,其次为化肥施用导致的温室气体排放,占比为21.18%。2007—2017年吉林省水稻生产的水足迹呈波动下降趋势,年均值为147 L·kg^-1,其中:水稀缺足迹为122 L·kg^-1,约占83%,水劣化足迹为25 L·kg^-1,约占17%。碳足迹和水足迹的高值区和低值区在空间上分布不一致,吉林省西部和中部地区的水足迹较大、碳足迹较小,而东部、中东部地区碳足迹较大、水足迹较小。相关分析表明,碳足迹和水足迹呈负相关。     

A virulence locus of Pseudomonas aeruginosa encodes a protein secretion apparatus

Bacterial pathogens frequently use protein secretion to mediate interactions with their hosts. Here we found that a virulence locus (HSI-I) of Pseudomonas aeruginosa encodes a protein secretion apparatus. The apparatus assembled in discrete subcellular locations and exported Hcp1, a hexameric protein that forms rings with a 40 angstrom internal diameter. Regulatory patterns of HSI-I suggested that the apparatus functions during chronic infections. We detected Hcp1 in pulmonary secretions of cystic fibrosis (CF) patients and Hcp1-specific antibodies in their sera. Thus, HSI-I likely contributes to the pathogenesis of P. aeruginosa in CF patients. HSI-I-related loci are widely distributed among bacterial pathogens and may play a general role in mediating host interactions.     

闵行地区PRRSV的分离鉴定及N基因序列分析

采集闵行地区规模猪场疑似PRRS阳性病料,在Marc-145细胞上进行病毒分离,采用RT-PCR扩增PRRSV高度保守的N基因并测序。结果表明,成功分离到6株PRRSV,为研究闵行地区PRRSV流行株分子变异和致病机理提供了良好的生物素材。