ADCY2基因表达对延边黄牛脂肪前体细胞成脂分化的影响

本试验旨在研究环腺苷酸（3',5'-cyclic adenosine monophosphote,cAMP）环化酶合成酶2（adenylyl cyclase 2,ADCY2）基因对延边黄牛脂肪前体细胞成脂分化的影响。选取3日龄的延边黄牛腹股沟皮下脂肪组织进行脂肪前体细胞的分离、培养和诱导分化;设计ADCY2基因的干扰片段（siADCY2-1、2、3）,构建pEX4过表达载体;分别收集正常诱导（对照组）、干扰和过表达0、5和10 d的脂肪细胞。用实时荧光定量PCR检测过氧化物酶体增殖激活物受体（PPARγ）、CCAAT/增强子结合蛋白（C/EBPα）和ADCY2在细胞分化过程中mRNA的转录水平,并用Western blotting法检测其蛋白的表达;用油红O染色检测脂滴含量的变化;用甘油三酯试剂盒检测甘油三酯含量。试验结果表明,在成脂分化过程中,与未分化相比,ADCY2基因在分化的第5和10天表达量均极显著上升（P<0.01）,而分化第10天与第5天相比水平极显著下降（P<0.01）;siADCY2-1干扰效率最高,过表达ADCY2基因使其mRNA水平升高;与对照组相比,过表达组细胞内ADCY2基因mRNA水平提高了约4 000 000倍,脂肪细胞内脂滴含量和甘油三酯的含量极显著提高（P<0.01）,成脂关键基因C/EBPα和PPARγ表达极显著上调（P<0.01）,而RNA干扰组细胞内ADCY2基因mRNA水平极显著降低（P<0.01）,脂肪细胞内脂滴含量和甘油三酯的含量极显著下降,成脂关键基因C/EBPα和PPARγ表达极显著下调（P<0.01）。因此,ADCY2基因过表达能显著增加成熟脂肪细胞的脂滴含量及甘油三酯含量,促进成脂关键基因C/EBPα和PPARγ的表达,说明ADCY2基因对脂肪细胞分化具有正向调控作用。     

miR-17-3p Regulates Preadipocyte Differentiation by Targeting KCTD15 in Yanbian Yellow Cattle

The aim of this study was to explore whether miR-17-3p could target KCTD15 to regulate the preadipocytes differentiation of Yanbian Yellow cattle. Bioinformatics softwares were used to identify homology and predict target genes of miR-17-3p. miR-17-3p mimic or miR-17-3p inhibitor and their negative control were transfected into bovine preadipocytes to overexpress or inhibit the expression of miR-17-3p. The binding sites of miR-17-3p to KCTD15 were verified via double luciferase report system. qRT-PCR and Western blot were used to detect the effect of miR-17-3p on the expressions of PPARγ, C/EBPα and KCTD15 both at mRNA and protein levels. Bioinformatics software prediction showed that KCTD15 was the target gene of miR-17-3p, and miR-17-3p had high homology in mammals. The mRNA and protein expressions of adipogenic marker genes PPARγ and C/EBPα after transfection of miR-17-3p mimic were significantly or extremely significantly higher than those of the control group transfected with NC-mimic (P<0.05 or P<0.01). After the expression of miR-17-3p was inhibited, the expressions of PPARγ and C/EBPα were significantly or extremely significantly lower than that in the control group (P<0.05 or P<0.01). Through the dual-luciferase reporter assay verification, the overexpression of miR-17-3p extremely significantly inhibited the fluorescent activity of the luciferase reporter gene vector containing the 3'UTR fragment of KCTD15 (P<0.01). Overexpression of miR-17-3p could also significantly or extremely significantly inhibit the expression of KCTD15 mRNA and protein(P<0.05 or P<0.01). However, inhibition of miR-17-3p expression could significantly increase the expression of KCTD15 mRNA and protein(P<0.05). These results suggest that miR-17-3p may regulate the differentiation of preadipocytes in Yanbian Yellow cattle by inhibiting the expression of KCTD15.     

miR-17-3p靶向KCTD15调控延边黄牛前体脂肪细胞分化

旨在探究miR-17-3p是否可以通过靶向KCTD15调节延边黄牛前体脂肪细胞的分化。本研究使用生物信息学软件鉴定miR-17-3p的同源性,预测miR-17-3p的靶基因;通过在延边黄牛的前体脂肪细胞中转染miR-17-3p mimic或miR-17-3p inhibitor及阴性对照实现细胞内miR-17-3p过表达或抑制表达;采用双荧光素酶报告基因检测系统确定miR-17-3p与KCTD15的靶标关系;使用qRT-PCR和Western blot技术在mRNA和蛋白水平上检测miR-17-3p对KCTD15基因以及脂代谢标志基因PPARγ和C/EBPα表达水平的影响。结果表明,生物信息学软件预测KCTD15可作为miR-17-3p的靶基因,且miR-17-3p在哺乳动物中具有较高的同源性;转染miR-17-3p mimic后脂代谢标志基因PPARγ和C/EBPα的mRNA和蛋白表达量显著或极显著高于转染NC-mimic的对照组（P<0.05或P<0.01）;抑制miR-17-3p的表达后,PPARγ和C/EBPα的表达量则显著或极显著低于对照组（P<0.05或P<0.01）;双荧光素酶报告基因验证分析显示,过表达miR-17-3p极显著抑制了含有KCTD15 3'UTR片段的载体的荧光活性（P<0.01）;过表达miR-17-3p也会显著或极显著抑制候选靶基因KCTD15 mRNA和蛋白质的表达量（P<0.05或P<0.01）;而抑制miR-17-3p的表达则会显著提高靶基因KCTD15的mRNA和蛋白质的表达量（P<0.05）。这些结果说明,miR-17-3p可能通过抑制KCTD15的表达促进延边黄牛前体脂肪细胞分化。     

Polymorphism of FGF14 Gene and Its Effect on Growth Traits in Yanbian Yellow Cattle

The purpose of this study was to explore the polymorphism of exon 3 of fibroblast growth factor 14 (FGF14) gene and its effect on growth traits in Yanbian Yellow cattle.The third exon of FGF14 gene was sequenced and analyzed by DNA direct sequencing method,and the genotyping of FGF14 gene was detected by HRM typing technique with high resolution melting curve,so as to explore the relationship between different genotypes of FGF14 gene and growth traits of 16 and 24 months of age in Yanbian Yellow cattle.The results showed that one A>G mutation was detected at position 631 947 bp of FGF14 gene,two alleles of A and G,and three genotypes of AA,AG,and GG.The results of χ² test showed that the distribution of genotype frequency and allele frequency of FGF14 gene in two Yanbian Yellow cattle populations of 16 and 24 months of age conformed to Hardy-Weinberg equilibrium (P>0.05).Polymorphic information content (PIC) analysis results showed that FGF14 gene was moderately polymorphic in both groups (0.25 < PIC < 0.5).The results of association analysis with growth traits showed that the different genotypes of FGF14 gene had significant influence on the traits of height at back and rump length of Yanbian Yellow cattle at 16 months of age (P<0.05),and had significant influence on the traits of body height,body weight and height at hip cross of Yanbian Yellow cattle at 24 months of age (P<0.05).In conclusion,it was speculated that FGF14 gene had a certain influence on the growth and development of Yanbian Yellow cattle,and provided reference for the subsequent modern molecular breeding and breeding of meat traits in Yanbian Yellow cattle.     

延边黄牛FGF14基因多态性及其对生长性状的影响

本研究旨在探索延边黄牛成纤维细胞生长因子14（fibroblast growth factor 14,FGF14）第3外显子多态性及其对生长性状的影响。应用DNA直接测序法对延边黄牛FGF14基因第3外显子进行测序分析,利用高分辨率熔解曲线HRM分型技术对FGF14基因进行分型检测,探究FGF14基因不同基因型与16和24月龄延边黄牛生长性状的关联性,并进行遗传统计学分析。结果显示,在FGF14基因的631 947 bp处检测到1处A>G突变,有2个等位基因：A和G,存在3种基因型：AA、AG和GG。χ²检验结果表明,FGF14基因的基因型频率和等位基因频率在16和24月龄延边黄牛群体中的分布均符合Hardy-Weinberg平衡定律（P>0.05）。多态信息含量（PIC）分析表明,FGF14基因在这2个群体中均处于中度多态（0.25 < PIC < 0.5）。与生长性状关联分析结果显示,FGF14基因不同基因型对16月龄延边黄牛背高和尻长均具有显著影响（P<0.05）,对24月龄延边黄牛体高、体重、十字部高均具有显著影响（P<0.05）。综上所述,推测FGF14基因对延边黄牛生长发育有一定影响,为后续延边黄牛肉质性状的现代分子育种选育工作提供参考。     

延边黄牛DGATs基因克隆、生物信息学及组织表达分析

试验旨在克隆延边黄牛二酯酰甘油酰基转移酶（diacylglycerol acyltransferase,DGATs）两种亚型（DGAT1和DGAT2）的CDS区核苷酸序列,并根据生物信息学对两种亚型的氨基酸序列进行分析,以及探讨其在延边黄牛各组织中的表达规律。采用RT-PCR和实时荧光定量PCR技术分别进行DGATs基因CDS区的扩增、克隆以及其在延边黄牛8个组织中mRNA表达量的检测,利用NCBI中BLAST将其与其他物种进行相似性分析,并构建系统进化树;通过在线工具对其编码蛋白的理化性质、一级结构及高级结构进行预测。结果显示,DGAT1和DGAT2基因CDS区序列长度分别为1 470和1 086 bp,分别编码489和361个氨基酸;二者均为稳定的疏水性蛋白。DGAT1存在25个潜在的磷酸化位点、1个N-糖基化位点和8个跨膜结构域;DGAT2存在28个潜在的磷酸化位点、2个N-糖基化位点和1个跨膜结构域。二者均不存在信号肽,即不属于分泌蛋白。DGAT1主要是通过α-螺旋和无规则卷曲连接,而DGAT2以α-螺旋、无规则卷曲和延伸链连接为主。延边黄牛DGAT1和DGAT2基因分别在延边黄牛小肠和脂肪组织中表达量最高,且显著高于其他组织（P<0.05）。本试验结果对进一步研究延边黄牛DGATs基因以及探究其在脂肪沉积过程中的作用机制具有重要意义。     

延边黄牛ACTA1基因的SNPs检测及其与肉质性状的关联分析

试验旨在研究延边黄牛ACTA1基因的遗传变异,分析其多态性对肉质性状的影响。采用PCR-RFLP和PCR-SSCP方法探讨ACTA1基因多态性及其与肉质性状的关联性。结果发现,延边黄牛ACTA1基因存在2个突变位点:A193G突变,导致氨基酸Arg → Gly的错义突变;A298G突变,导致氨基酸Lys →Arg的错义突变。χ²检验显示,A193G和A298G位点于检测群体中处于Hardy-Weinberg平衡状态（P>0.05）。关联分析显示,延边黄牛ACTA1基因2个突变位点的不同基因型与肌肉中肉豆蔻酸、油酸含量、眼肌面积、大理石花纹等级呈显著相关（P<0.05）。初步认为ACTA1基因对延边黄牛肉质性状有显著影响,可作为延边黄牛新品种早期选择的候选基因。     

Polymorphism of ACTA1 Gene and Its Association with Meat Quality Traits in Yanbian Yellow Cattle

This study was aimed to research the gentic variations of ACTA1 gene in Yanbian Yellow cattle, and analyze the influence of genetic polymorphisms on meat quality traits. PCR-RFLP and PCR-SSCP technique were used to analyze the association between the polymorphisms of ACTA1 gene and meat quality traits. The results showed that there were two SNPs, A193G which with replacement of Arg by Gly amino acid, A298G which with replacement of Lys by Arg amino acid. χ² test showed A193G and A298G were in Hardy-Weinberg equilibrium (P>0.05). The association analysis indicated that the genotypes of different mutations in ACTA1 gene were found to be significantly related to myristic acid, oleic acid, loin muscle area, marbling score (P<0.05). ACTA1 gene had potential effects on meat quality traits of Yanbian Yellow cattle, and could be used for candidate gene in early selection of new varieties of Yanbian Yellow cattle.     

延边牛PLIN2基因CDS区克隆、生物信息学分析及其表达规律研究

本研究旨在克隆延边牛脂肪分化相关蛋白PLIN2基因完整CDS区,通过生物信息学分析PLIN2基因CDS区序列和蛋白质基本特性,探讨其在延边牛不同组织及前体脂肪细胞成脂过程中的表达规律。选取18月龄去势的延边牛为研究对象,利用RT-PCR和基因克隆获得延边牛PLIN2基因,与其他物种进行同源性比对及系统进化树构建,利用生物信息学方法预测PLIN2编码蛋白质的理化性质、潜在磷酸化位点及糖基化位点、二硫键分析、信号肽、亚细胞定位、蛋白高级结构,利用实时荧光定量PCR检测PLIN2基因在延边牛不同组织中的表达水平。结果显示,延边牛PLIN2基因CDS长1 353 bp,编码450个氨基酸;同源性比对发现延边牛PLIN2基因与黄牛、水牛、绵羊、山羊、野猪、人、小鼠和野鸡的同源性分别为100%、98.7%、96.7%、97.1%、85.5%、86.3%、47.3%和49.1%;系统进化树表明,延边牛与黄牛、水牛的亲缘关系最近,与野鸡的亲缘关系最远。PLIN2蛋白为不稳定蛋白,具有一定亲水性,存在61个潜在的磷酸化位点、16个O-糖基化潜在位点、12个N-糖基化潜在位点,存在2个二硫键,不存在信号肽,主要分布于细胞核中,细胞质和线粒体中有少量分布。PLIN2蛋白高级结构预测该蛋白是由α-螺旋、延伸链、无规则卷曲和β-转角组成,为混合型蛋白,通过无规则卷曲连接,以α-螺旋为主。实时荧光定量PCR结果显示,PLIN2基因在延边牛小肠组织中表达量最高,其次为背最长肌和肾脏组织。本研究结果可为进一步研究PLIN2基因的功能提供借鉴。     

延边黄牛UCP2、UCP3基因多态性及其与生长性状的关联分析

试验旨在探究延边黄牛解耦联蛋白2（uncoupling protein 2,UCP2）和解耦联蛋白3（uncoupling protein 3,UCP3）基因多态性及其与生长性状的相关性。以120头24月龄的延边黄牛为试验对象,提取血液基因组DNA后构建混池,运用DNA测序法对UCP2、UCP3基因进行测序分析,应用高分辨率熔解曲线分型技术对延边黄牛UCP2、UCP3基因进行分型检测,并结合生长性状数据进行关联分析。结果显示,在24月龄延边黄牛群体中,UCP2基因53 412 568 bp处存在C>G突变（g.53412568 C>G）,产生2个等位基因：C和G,形成3种基因型：CG （0.358）、GG （0.125）和CC （0.516）;UCP3基因53 443 536 bp处存在C>T突变（g.53443536 C>T）,产生2个等位基因：C和T,形成3种基因型：CT （0.383）、CC （0.450）和TT （0.167）。关联分析显示,UCP2基因g.53412568 C>G位点CC基因型体重、腰角宽均显著高于GG基因型,CG基因型个体背高显著高于GG基因型（P<0.05）;UCP3基因g.53443536 C>T位点CC基因型胸围显著高于TT基因型（P<0.05）。以上结果表明,UCP2和UCP3基因在延边黄牛生长发育过程中起到一定作用,可为延边黄牛现代分子育种后续研究提供参考。     

延边黄牛α-肌动蛋白1基因的多态性及其与生长性状的相关分析

试验旨在寻找肌内脂肪含量差异极显著的两组个体延边黄牛背最长肌组织差异表达基因,并分析其遗传多态性对生长性状的影响。应用引物退火技术获得了在肌内脂肪含量差异极显著的两组个体背最长肌组织差异表达α-肌动蛋白1(actin alpha 1,ACTA1)基因及其全长cDNA序列,该基因主要在心脏和背最长肌中表达。序列比对结果发现,在5'端非编码区193 bp处存在1个突变。采用PCR-RFLP方法对100头延边黄牛群体进行检测,发现AA、AG、GG 3种基因型,基因型频率分别为0.24、0.50、0.26;A、G等位基因频率分别为0.37、0.63。对该突变位点的多态性与生长性状的关联性进行分析,结果发现,在体长、体重和平均日增重上,GG基因型个体显著高于AA和AG基因型个体(P＜0.05);在胸围上,GG基因型个体显著高于AG基因型个体,但与AA基因型个体差异不显著(P＞0.05);在体高上,不同基因型之间差异不显著(P＞0.05)。初步认为GG基因型是提高延边黄牛体质量和体尺指标性状的有利基因型,提示ACTA1基因有可能作为延边黄牛生长性状的候选基因。     

Polymorphism of PLIN Gene and Its Associaton with Carcass and Meat Quality Traits in Yanbian Yellow Cattle

This experiment was conducted to explore the association between perilipin (PLIN) gene and carcass and meat quality traits in Yanbian Yellow cattle.In this study,70 30-month-old healthy Yanbian Yellow cattle were used as experimental materials,the blood of jugular vein and its 12-13 intercostal longissimus muscle dorsi were collected to extract DNA samples of Yanbian Yellow cattle,and determine the carcass and meat quality characteristics.The SNP of PLIN gene was detected by direct sequencing,and the correlation analysis was conducted based on the carcass and meat quality data of Yanbian Yellow cattle and the genetic polymorphism of PLIN gene.The results showed that there were 2 SNPs of PLIN gene exon 3 in Yanbian Yellow cattle:g.103 C→T and g.156 T→C,both of them were missense mutations.g.103 C→T contained two genotypes:CC and CT,CC genotype was the dominant genotype,the genotype frequency was 0.81,C was the dominant allele,the gene frequency was 0.90.The intramuscular fat,pre slaughter weight,carcass weight and back fat thickness of CT genotype was significantly higher than those of CC genotype(P<0.05).g.156 T→C contained two genotypes:TT and TC,TT genotype was the dominant genotype,the genotype frequency was 0.83,T was the dominant allele,the gene frequency was 0.92.The pre slaughter weight,carcass weight and back fat thickness of TC genotype were significantly higher than those of TT genotype (P<0.05).Chi square test showed that the two SNPs of PLIN gene in Yanbian Yellow cattle reached Hardy-Weinberg equilibrium (P>0.05),and belonged to low polymorphism (PIC<0.25).In conclusion,g.103 C→T and g.156 T→C of PLIN gene had potential effect on carcass and meat quality traits in Yanbian Yellow cattle,and could be used as a candidate gene and potential molecular marker of Yanbian Yellow cattle.     

延边黄牛PLIN基因多态性及其与胴体和肉质性状的相关性分析

为探究延边黄牛脂滴包被蛋白（perilipin,PLIN）基因遗传多态性及其对胴体和肉质性状的影响,试验选取70头30月龄健康延边黄牛公牛,采集颈静脉血液及其12~13肋间背最长肌以提取延边黄牛DNA样本,并测定胴体与肉质性状,采用直接测序方法检测PLIN基因SNP位点,结合延边黄牛胴体、肉质数据与PLIN基因遗传多态性进行关联分析。结果显示,在延边黄牛PLIN基因外显子3 103与156 bp处存在2个突变位点：g.103 C→T和g.156 T→C,均为错义突变。g.103 C→T位点存在CC、CT两种基因型,CC基因型为优势基因型（0.81）,C基因为优势等位基因（0.90）,表现为CT基因型个体脂肪含量、宰前重、胴体重、背膘厚均显著高于CC基因型个体（P<0.05）;g.156 T→C位点存在TC、TT两种基因型,TT基因型为优势基因型（0.83）,T基因为优势等位基因（0.92）,表现为TC基因型个体的宰前重、胴体重及背膘厚均显著高于TT基因型个体（P<0.05）。卡方检验结果表明,延边黄牛在PLIN基因外显子3的2个SNPs位点均达到Hardy-Weinberg平衡（P>0.05）,杂合度较低,属于低度多态（PIC<0.25）。综上,PLIN基因g.103 C→T和g.156 T→C位点对延边黄牛部分胴体与肉质性状存在一定影响,该位点可作为延边黄牛胴体、肉质性状的潜在标记,PLIN基因可以作为延边黄牛品种选育的候选基因和潜在分子标记。     

miR-93对延边黄牛垂体细胞GH分泌的影响

为了分析血液外泌体miRNA对延边黄牛垂体细胞生长激素（GH）分泌的影响,试验选择在延边黄牛和韩延牛血液外泌体中存在显著差异表达的miR-93,分析其对延边黄牛垂体细胞GH分泌的影响机制。试验首先进行延边黄牛垂体细胞的原代培养,之后将miR-93的mimics（miR-93-mi组）、mimics对照品（NC对照组）、inhibitor（miR-93-in组）、inhibitor对照品（iNC组）转染给已建立的垂体原代细胞,48 h后收集细胞,提取总mRNA和总蛋白。试验利用targetscan和RNAhybrid分析软件对miR-93的靶基因进行预测,并利用双荧光素酶报告基因系统对miR-93的靶关系进行验证;利用实时荧光定量PCR和Western blotting技术分别检测靶基因mRNA转录和蛋白的表达情况,结果表明,miR-93靶向了生长激素释放激素受体（GHRHR）的3'UTR;与NC对照组比较,miR-93-mi组的延边黄牛垂体细胞中GH mRNA转录和蛋白表达均极显著低于NC对照组（P<0.01）;miR-93-mi组的GHRH mRNA转录和蛋白表达均极显著低于NC对照组（P<0.01）,而miR-93-in组的GHRHR蛋白表达显著高于iNC对照组（P<0.05）。说明miR-93可通过调节GHRHR的表达而调控延边黄牛垂体细胞GH的分泌,进而调控延边黄牛的生长发育。     

The Correlation Analysis Between the SNPs of MC4R Gene and Meat Quality Traits in Yanbian Yellow Cattle

In order to explore the effects of the key gene of regulating cattle fat deposition on meat quality traits, screening candidate gene which affect beef fattening traits. This study employed 157 cattles (24 months old) with similar weight as research object, analyzed the correlation between SNPs of MC4R gene and meat quality traits in Yanbian Yellow cattle by PCR direct sequencing method. The results indicated that the mutation site of C→G had been found in MC4R gene 1 069 bp, which resulted in L (leucine) translated into V (valine). Protein secondary structure lost α-spiral and β-fold at the 166, 243 and 280 amino acid position, which resulted in the changes of spatial position of β-turns in tertiary structure. This site showed different degrees correlation with backfat thickness, lrightness, yellowness of Yanbian Yellow cattle. The backfat thickness and yellowness (3 d) of GG genotype were significantly higher than CC and CG genotypes (P <0.05). The lrightness of CC and CG genotypes were significantly higher than CC genotype (P <0.05). These results declared that the site had remarkably correlatived with Yanbian Yellow cattle relevant economic traits, and the mutation site could be used as a potential molecular marker.     

延边黄牛黑皮质素受体-4基因多态性及其与肉质性状的相关性分析

试验旨在探索调控肉牛脂肪沉积的关键基因对肉质性状的影响,筛选影响肉牛育肥性状的候选基因。本研究以157头24月龄、体重相近的延边黄牛为研究对象,利用PCR扩增直接测序的方法,分析了延边黄牛黑皮质素受体-4（melanocortin-4 receptor,MC4R）基因的SNPs及其与肉质性状的相关性。结果发现,延边黄牛MC4R基因在1 069 bp处存在C→G突变,导致L （亮氨酸）转变为V （缬氨酸）,引起蛋白质二级结构在166、243、280位氨基酸处出现α螺旋、β折叠丢失,导致三级结构中β转角的空间位置改变。经关联分析表明,该位点与延边黄牛背膘厚、亮度、黄度都存在不同程度的相关,其中GG基因型背膘厚与屠宰后黄度（3 d）显著高于CC和CG基因型（P＜0.05）;GG、CG基因型屠宰后亮度显著高于CC基因型（P＜0.05）。说明该位点与延边黄牛肉质性状存在显著相关,该突变位点可作为潜在的分子标记。     

海藻糖替代部分甘油对延边黄牛冻融精子质量的影响

【目的】探讨是否可以通过补充海藻糖来降低冷冻保护剂中甘油的浓度,从而提高冻融精子的质量。【方法】分别用6%甘油(Ⅰ组)及3%甘油+0、50、100和150 mmol/L海藻糖(Ⅱ、Ⅲ、Ⅳ和Ⅴ组)处理精子,用精子-微生物动(静)态图像检测分析系统(CASA)检测冻融精子的活力、质膜完整性和顶体完整性及动力学相关参数,筛选出最佳海藻糖处理浓度用于后续试验。通过Western blotting方法检测精子蛋白酪氨酸磷酸化水平评价精子获能状态,Hoechst 33342/PI/JC-1联合染色法检测精子的活率及线粒体膜电位,色霉素A₃(CMA₃)染色法检测精子DNA的完整性,部花青540(M540)和Yo-Pro-1染色检测精子膜脂质紊乱水平。【结果】与Ⅰ组相比,Ⅱ组精子的活力、质膜完整性、顶体完整性以及曲线速度(VCL)和直线率(STR)均显著下降(P<0.05),Ⅲ、Ⅳ和Ⅴ组组精子的活力、VCL、直线速度(VSL)、平均路径速度(VAP)、直线性运动(LIN)和STR均显著升高(P<0.05)。因此,后续试验选用100 mmol/L海...     

延边黄牛体细胞核移植的研究进展

牛体细胞核移植(somatic cell nuclear transfer,SCNT)是一套极其复杂的技术体系,它包括卵母细胞的成熟、供核细胞的准备、卵母细胞的显微操作、细胞融合、卵母细胞激活和胚胎培养。因此,许多因素影响着核移植胚胎发育。虽然陆续有克隆牛出生的报道,但是克隆效率依然低下。本综述对延边黄牛体细胞核移植体系的6个方面进行简单综述,以期为探究延边黄牛体细胞克隆的最优化条件、建立最优化的培养体系、提高克隆效率提供参考。     

Research Advances on Somatic Cell Nuclear Transfer in Yanbian Yellow Cattle

Bovine somatic cell nuclear transfer (SCNT) is a sophisticated technique system,including oocyte maturation,donor cell preparation and oocytes microscopic operation,fusion,activation and culture.Although the birth of cloning cattle has been reported recently,the efficiency of somatic cell cloning has remained lowly.In order to establish the optimization somatic cell nuclear transfer system of Yanbian Yellow cattle,this review summarized only from 6 main aspects mentioned above in this field.