莱芜猪合成系的培育

随着现代养猪生产的发展和人民生活水平的不断提高 ,对瘦肉的需求量越来越大 ,高质量的瘦肉型猪倍受消费者的欢迎。莱芜猪是莱芜市当家的猪种 ,被誉为“国宝” ,以适应性强、繁殖率高、杂交优势明显 ,肉质细嫩香醇等优点而著称。自“八五”期间实施“莱芜猪自群选育及杂交利用研究”项目以来 ,莱芜猪经过 4个世代的保纯选育 ,生产性能得以巩固提高 ,并通过引入国外瘦肉型种公猪进行二元、三元杂交 ,在发展瘦肉猪生产中起到了积极的作用。特别是大莱二元母猪 ,繁殖性能突出 ,其他性能也较好。但进入 90年代以来 ,随着我国规模化、工厂化养猪…     

影响奶牛体细胞手工克隆技术因素的研究

旨在优化奶牛的手工体细胞克隆技术方案,本试验以牛卵母细胞为材料研究了2个半卵不同粘合交流电压、重构胚不同激活方法和不同的COCs采集方法对奶牛手工体细胞克隆胚发育的影响.结果表明,(1)降低2个半卵粘合交流电压能显著提高其随后的卵母细胞融合率(P<0.05);(2)A23187+6-DMAP组激活的无透明带重构胚分裂率和囊胚率显著高于Ionomycin+6-DMAP组(P<0.05);(3)真空蠕动泵抽吸法和刀片切割法获得的卵母细胞总数显著高于注射器抽吸法(P<0.05),而切割法获得的1和2级卵母细胞百分数显著高于其它2种方法(P<0.05).因此,采用真空蠕动泵抽吸法获得COCs,用较低的粘合交流电压进行半卵粘合,采用A23187+6-DMAP激活法进行融合后的激活能显著提高奶牛手工体细胞克隆效率.     

哺乳动物体细胞手工克隆研究进展

采用化学去核和无透明带技术相结合,对哺乳动物卵母细胞进行处理,使通过该方法获得的去核无透明带卵母细胞作为受体,进行体细胞核移植,从而获得克隆动物。该方法完全避免了传统核移植的显微操作及其繁琐程序,是纯粹的手工克隆,它的成功将会大大简化核移植程序,提高核移植总效率,同时提高核移植的生产力。论文对手工克隆的研究进展,手工克隆的机理,手工操作的体细胞核移植方法及影响手工克隆的因素进行综述。     

莱芜猪的选育

莱芜猪是我国华北型猪中黄淮海黑猪的一个类群 ,是山东有代表性的一个地方良种 ,中心产区在莱芜市 ,素以适应性强、繁殖力高、肉质优良而著称。在漫长历史进程中形成的这个古老的地方猪种 ,过去从未系统地进行过选育。建国后 ,在大规模的猪种改良过程中 ,曾一度濒临灭绝的边缘。 2 0世纪 70年代中期 ,山东投资兴建莱芜猪育种繁殖场 ,从社会上选取优秀个体组成保种群。 80年代中期开始有计划的进行品系繁育 ,现已完成 7个世代。 2 0多年来 ,坚持保种与开发相结合、以选育促利用、以利用保选育的基本指导思想 ,选育工作取得显著成效。目前 ,莱…     

莱芜猪胴体切块质量预判

本文以莱芜猪肉质育种为例,介绍了在屠宰线上对胴体的肉色、大理石纹、特别是肌内脂肪进行主观评定预测的简单易行的方法,其结果可用于肉质选种选育。     

莱芜猪种质资源调查报告

莱芜猪被称为华北第一猪,是我国国宝级地方良种。本文通过对莱芜猪资源的全面调查,了解其存在数量、分布区域、体貌特征、形成条件、各项性能等近况,为莱芜猪的文化挖掘、保护、开发、利用打下坚实基础。     

莱芜猪母猪生殖器官发育测定报告

引言莱芜猪的高繁殖力特性已为世人瞩目。据84－89年三个世代育种猪群251头（次）繁殖母猪测定，窝平均产仔14．76头，产活仔12．35头，其中窝产仔15头以上个体占群体30％。最高产仔28头，活仔22头．群众反映小母猪生后3月龄即有发情表现，此间输配的小猪可以怀孕。为研究莱芜母猪的住早熟期，按排本试验。本研究目的：①了解母猪生殖器官发育变化特点；②卵泡发育及排卵期。为研究莱芜猪多胎高产的特性提供理论依据。本试验93年24／4起到7／8，在莱芜市杨庄莱芜猪育种繁殖场进行。材料与方法①于莱芜母猪30、40、50、60……120日龄阶段测定…     

莱芜猪种质资源的保护利用研究与开发

改革开放30多年,通过制定正确的发展方案和保种选育技术路线,对莱芜猪进行了9个世代的保纯选育;同步对莱芜猪的肉质、繁殖、营养需求、生长发育和杂交配合力进行了深入研究。并利用莱芜猪创新性地培育出了鲁莱黑猪、鲁农I号猪配套系、欧得莱猪配套系、莱芜猪合成系、大莱、汉大莱、杜长大莱优秀瘦肉型猪等新种质;在保种、研究、利用的同时,对其培育的新种质进行了社会性的推广、示范及产业开发,取得了巨大的社会经济效益。莱芜猪已成为全国许多地区、单位生产与育种的首选种质材料,是生产优质特色品牌猪肉最优良的种质资源。     

猪克隆技术的研究进展

自从1986年我国首次自主成功克隆猪,克隆技术在猪的研究获得较大推进,国内陆续报道了地方品种猪的成功克隆.然而,对于克隆猪在生产中的应用研究,暂时未见有较多报道.本文从动物克隆技术的发展、国内外研究进展以及克隆技术在生猪生产应用中存在的难点和缺点进行阐述,进一步推动猪克隆技术在猪生产中应用.     

Comparison of efficiency of in vitro cloned sheep embryo production by conventional somatic cell nuclear transfer and handmade cloning technique

Conventional somatic cell nuclear transfer (SCNT) technique of in vitro production of cloned embryos involves use of costly and complicated micromanipulators. Handmade cloning (HMC) technique has been applied as efficient and cost‐effective alternative in many livestock species. The aim of the present study was to compare the efficiency of in vitro production and in vitro development of cloned sheep embryos by the two techniques. Cloned embryos were produced by conventional SCNT using micromanipulator apparatus and by HMC technique. Enucleation efficiency and efficiency of fusion with somatic cell (nucleus donor) were compared. Cleavage percentage was observed on day 2 of in vitro culture (IVC), and morula and blastocyst percentages were calculated on day 7 of IVC. Higher enucleation efficiency (96.98 ± 1.01 vs. 93.62 ± 1.03; p > .05) as well as fusion efficiency was obtained with HMC technique than with conventional SCNT (96.26 ± 1.34 vs. 92.63 ± 0.70, p < .05); 181 cloned sheep embryos were produced in vitro by conventional SCNT and 92 by HMC. Cleavage percentage observed on day 2 of in vitro culture was higher in HMC than SCNT (66.92 ± 3.72 vs. 55.97 ± 2.5, respectively, p < .05). Morula percentage obtained was higher in SCNT than HMC (44.12 ± 2.93 vs. 30.43 ± 6.79, respectively, p < .05), whereas blastocyst percentage obtained by HMC was higher (12.46 ± 4.96) than SCNT (5.31 ± 2.25; p > .05). It was inferred that HMC technique provides a cost‐effective and efficient method of in vitro production of cloned sheep embryos with a comparatively simpler technique with a possibility of automation. Efficiency of cloned embryo production could be improved further by propagating and standardizing this technique.     

生长育肥猪的饲养管理

猪的生长育肥阶段是指小猪体重达到30公斤后，在被运往屠宰场前的最后一个阶段。饲养育肥猪的丰要目标是猪只生长速度快、饲料利用率高、屠宰品质好、饲养周期短及病死率低。实际生产中，生长育肥猪被统一饲养在一个固定区域，所以，要充分认识该区域对猪只的影响。     

Study of the efficiency of chemically assisted enucleation method for handmade cloning in goat (Capra hircus)

The present investigation was carried out to find an efficient chemically assisted procedure for enucleation of goat oocytes related to handmade cloning (HMC) technique. After 22-h in vitro maturation, oocytes were incubated with 0.5 μg/ml demecolcine for 2 h. Cumulus cells were removed by pipetting and vortexing in 0.5 mg/ml hyaluronidase, and zona pellucida were digested with pronase. Oocytes with extrusion cones were subjected to oriented bisection. One-third of the cytoplasm with the extrusion cone was removed with a micro blade. The remaining cytoplasts were used as recipients in HMC. Goat foetal fibroblasts were used as nuclear donors. The overall efficiency measured as the number of cytoplasts obtained per total number of oocytes used was significantly (p < 0.05) higher in chemically assisted handmade enucleation (CAHE) than oriented handmade enucleation without demecolcine (OHE) (80.02 ± 1.292% vs. 72.9 ± 1.00%, respectively, mean ± SEM). The reconstructed and activated embryos were cultured in embryo development medium (EDM) for 7 days. Fusion, cleavage and blastocyst development rate were 71.63 ± 1.95%, 92.94 ± 0.91% and 23.78 ± 3.33% (mean ± SEM), respectively which did not differ significantly from those achieved with random handmade enucleation and OHE. In conclusion, chemically assisted enucleation is a highly efficient and reliable enucleation method for goat HMC which eliminates the need of expensive equipment (inverted fluorescence microscope) and potentially harmful chromatin staining and ultraviolet (UV) irradiation for cytoplast selection.     

Molecular cloning and expression analysis of pig CD7

CD7 is an integral membrane protein which mediates an important signal to mediate the differentiation, activation, and regulation of some T cells and NK cells. However, only human and mouse CD7 have been identified and studied among mammalian species. In this study, we cloned pig CD7 cDNA and determined its complete cDNA sequence. Pig CD7 cDNA contained an open reading frame (627 bp) encoding 208 amino acids with well conserved motifs involved in signal transduction within cytoplasmic tail among mammalian species. Pig CD7 mRNA was detected by RT-PCR in mainly lymphoid tissues, indicating the conserved functions of CD7 in pigs. Moreover, we generated soluble pig CD7 fusion immunoglobulin (pig CD7Ig) containing extracellular domain of pig CD7 to test whether pig CD7 binds to pig galectin-3. Flow cytometry and immunohistochemistry analyses indicated that soluble pig CD7Ig can bind to galectin-3 expressed in macrophages and epithelial cells of small intestine. These results help to analyze the structural relationship between CD7 and its ligand transferring signal transduction among mammalian species.     

Molecular cloning and expression analysis of pig CD81

CD81, also known as TAPA-1 (target of antiproliferative antibody 1), is a member of the tetraspanin family of proteins and a component of the B cell co-receptor complex. Several studies have shown that CD81 plays significant roles in a variety of immune responses, including activation of B cells and T cells. In this study, we cloned pig Cd81 cDNA using RT-PCR coupled with rapid amplification of cDNA ends (RACE)-PCR and determined the complete cDNA sequence of pig Cd81. Pig Cd81 cDNA contains an open reading frame (711 bp) encoding 236 amino acids. The identity of pig CD81 with those of human, cattle, rat, and mouse are 90.30%, 92.26%, 86.22%, and 86.22%, respectively. Alignment of the CD81 amino acid sequence with those of mammalian species showed that the large extracellular loop (LEL) is the most divergent, whereas other domains are largely conserved. Pig Cd81 mRNA was detected by RT-PCR in a broad range of tissues, including lymphoid tissues as well as nonlymphoid tissues, indicated variety of cellular functions of CD81 in most pig tissues. Flow cytometry analyses demonstrated that human CD81 antibody recognizes a pig CD81 on the cell surface. Further, immunohistochemistry analysis using human CD81 antibody on pig spleen was revealed that CD81 expression is widely diffused in spleen tissue. Future study will be focused on defining the functional role of CD81 during the course of pig infectious diseases.     

Advancing pig cloning technologies towards application in regenerative medicine

Regenerative medicine is expected to make a significant contribution by development of novel therapeutic treatments for intractable diseases and for improving the quality of life of patients. Many advances in regenerative medicine, including basic and translational research, have been developed and tested in experimental animals; pigs have played an important role in various aspects of this work. The value of pigs as a model species is being enhanced by the generation of specially designed animals through cloning and genetic modifications, enabling more sophisticated research to be performed and thus accelerating the clinical application of regenerative medicine. This article reviews the significant aspects of the creation and application of cloned and genetically modified pigs in regenerative medicine research and considers the possible future directions of the technology. We also discuss the importance of reproductive biology as an interface between basic science and clinical medicine.     

猪抗病育种研究进展

作者对开展抗病育种的必要性、发展过程、抗病的遗传基础、抗病育种方法的现状及存在的问题进行了介绍。其中对MHC、抗病主基因、抗病育种的直接、间接方法、转基因抗病育种进行了重点介绍，提出采用分子标记辅助选择和转基因工程抗病育种是当前和今后抗病育种的主要方向，最后对抗病育种将来的发展前景进行了展望。     

Molecular cloning and expression analysis of pig CD79alpha

The CD79alpha (immunoglobulin alpha, Igalpha), a part of B cell receptor (BCR) complex, forms a heterodimer with CD79beta (Igbeta) and plays an important role in the B cell signaling. In this study, we have cloned pig Cd79a cDNA using RT-PCR and determined the complete cDNA sequence of pig Cd79a. Pig Cd79a cDNA contains an open reading frame (672bp) encoding 223 amino acids. The putative amino acid identity of pig CD79alpha with those of human, cattle and mouse are 70.4, 81.4, and 67.7%, respectively. Alignment of the CD79alpha amino acid sequence with those of mammalian species showed that the extracellular domain is the most divergent, whereas transmembrane region and cytoplasmic tail including immunoreceptor tyrosine-based activation motif (ITAM) are largely conserved. Pig Cd79a mRNA was detected mainly in lymphoid tissues by RT-PCR. The highest level of Cd79a mRNA expression was observed in mesenteric lymph node and spleen. Relatively low level of Cd79a mRNA expression was observed in lung, thymus and small intestine. The lowest level of Cd79a mRNA expression was observed in large intestine. Flow cytometry analyses demonstrated that human CD79alpha antibody recognizes a CD79alpha in pig B cells. Further, immunohistochemistry analysis using human CD79alpha antibody on pig spleen was revealed that CD79alpha is strongly expressed in the follicular mantle zone rather than in the germinal center. Future study will be focused on defining the functional role of CD79alpha during the course of pig infectious diseases and the formation of neoplasm.     

猪CAPN3基因的克隆及PCR-SSCP分析

calpain3在肌肉组织中的特异表达,其活性显著影响宰后肉的嫩化。研究利用比较基因组学的方法首次克隆了猪CPAN3基因的内含子16~23,并利用PCR-SSCP方法在外显子1中寻找到1个插入/缺失突变和1个碱基替代,为进一步分析其变异和肉质的关系并进而确定遗传标记提供了基础。