微卫星标记与奶牛数量性状QTL定位

本文对奶牛产奶性能（产奶量、乳脂率、乳脂量、蛋白率、蛋白量）、乳房炎及体细胞数、繁殖性能、生产寿命等数量性状QTL定位研究进展进行了综述。 Abstract:Mapping of quantitative trait loci for some important traits (milk yield,fat percentage,fat yield,protein percentage,protein yield,clinical mastitis and somatic cell count,reproductive performance,productive life,etc.) in dairy cattle was introduced in this review.     

微卫星荧光标记基因组扫描

微卫星荧光标记基因组扫描为 2 0世纪 90年代发展起来的一项分子生物学技术 ,在基因定位、个体识别等方面有广泛的应用 .文中对其原理、策略、分析方法及应用作了论述 ,并探讨了它的发展前景 .     

微卫星标记及其在鸟类中的应用

微卫星DNA作为一种分子标记,以其诸多优点被认为是各类遗传标记中最有价值的一种。目前已被广泛应用于鸟类的研究中。本文介绍了微卫星DNA标记的分布特点、突变机制和作为分子标记的特点,并综述了微卫星DNA标记在鸟类的亲权分析及种类鉴定、遗传图谱构建及基因定位、物种进化历史揭示和遗传多样性研究中应用现状。     

衰老标记蛋白30 mRNA在癌细胞中的表达研究

目的:检测衰老标记蛋白(SMP)30 mRNA在不同癌细胞系中的表达情况,探讨其在不同细胞中的表达差异。方法:分别采用RT-PCR与荧光定量PCR检测SMP30 mRNA在正常肝细胞、肝癌细胞、胃癌细胞、乳腺癌细胞、宫颈癌细胞中的表达,并用SPSS13.0进行统计学分析。结果:SMP30 mRNA在所有被检测的细胞株中均有表达,在癌细胞中的相对表达量分别为肝癌细胞(0.926±0.340)、胃癌细胞(0.922±0.379)、乳腺癌细胞(0.614±0.356)、宫颈癌细胞(0.608±0.346),而在正常肝细胞中为0.175±0.158,显示SMP30 mRNA在癌细胞中的表达量较正常肝细胞中高(P0.05),且在肝癌细胞中的表达量比在其他癌细胞中更高。结论:SMP30 mRNA在癌细胞中的表达高于正常肝细胞,且在肝癌细胞中的表达高于其他癌细胞,具有临床应用价值。     

70个水稻微卫星标记染色体位置的更正

微卫星标记(SSR)因其操作简单和稳定可靠的特点而成为一种重要的分子标记,被广泛应用于遗传作图和种质鉴定等方面。但其在染色体上位置的正确性将直接影响到基因定位的正确性和后续研究的方向。利用美国国家生物信息技术中心(NCBI)网站的Blast程序,将2740个SSR标记的前后引物序列与水稻粳稻品种日本晴基因组进行比对,共发现70个标记位于另一条染色体,对这70个标记重新锚定的染色体进行了更正。这将有助于今后水稻分子标记遗传连锁图的正确构建。     

奶牛微卫星基因座与产奶性能关系的研究

根据小鼠与牛的遗传比较图谱及相关报道选择了与weaver基因连锁的7个微卫星基因库,并在荷斯坦牛群体中对这些基因座进行群体遗传学特性分析。选择具有中度以上多态性的4个微卫星基因座（BM6438、BMS2321、BMS711和TGLA116）进行产奶性能的相关分析。最小二乘分析结果表明,4个微卫星基因座对产奶量影响不显著（P＞0．05）,BM6438生BMS711基因座对乳成分影响不显著（P＞0．05）,BMS2321基因座对乳蛋白量和乳蛋白率有极显著的影响（P＜0．01）,TGLA116对乳蛋白量和乳蛋白率的影响达到0．05的显著水平。     

微卫星标记对资源猪群的遗传分析和连锁图谱构建

在猪数量性状位点的定位研究中,标记的使用和图谱的构建是很重要的。本研究从猪的第4、6、7、8和13染色体上选取39个微卫星标记,在来源于约克夏和梅山214头猪组成的资源群中,分析了遗传特征并构建了图谱。研究表明,平均等位基因数、平均观察杂合度(Ho)和平均多态信息含量(PIC)在F1和F2代中分别为:3.2,0.528,0.463和3.2,0.496,0.447。结果表明大多数微卫星标记位点表现为中高度杂合性。在资源群体中,平均有信息减数分裂数是217.4(44-316),而各染色体上两性平均图谱的长度分别是:172.3cM(SSC4),168.7cM(SSC6),191.7cM(SSC7),197.3cM(SSC8),178.3cM(SSC13)。与USDA-MARC的参考图谱相比,标记位点的顺序相同,但长度均较长。雌雄两性图谱相比,第4和第6染色体上雌性图谱长于雄性图谱;而在另外3条染色体上,则雄性图谱长于雌性图谱。结果显示了标记位点在资源猪群的遗传特征和遗传关系,其连锁图谱可用于今后的QTL定位。     

微卫星标记及其在动物遗传育种中的应用

微卫星标记技术一出现,就以其独特的优点引起了动物遗传育种学家的广泛关注,显示出较好的应用前景。本文就微卫星标记的特点及其在动物遗传育种中的应用作以综述。     

天麻基因组微卫星特征分析与分子标记开发

该研究基于第二代测序技术建立了天麻的基因文库,筛选微卫星序列,并对微卫星位点的类型、丰度、长度、偏好性等进行了分析与比较;并为60条重复次数高的微卫星序列设计了引物,运用4个种群80个样本进行了PCR扩增和聚丙烯酰胺凝胶电泳检测。结果表明:(1)天麻基因组测序得到61 048条基因序列,检测出微卫星位点12 107个,其中二核苷酸重复最多、长度变异大。(2)设计的60对微卫星引物中的20对能扩增出清晰条带且有多态性,每个位点的复等位基因数(N_a)在4~14之间,平均为8.40;多态性信息含量(PIC)平均为0.77。该研究开发的天麻微卫星分子标记为开展天麻遗传学研究及种质资源鉴定等工作奠定了基础。     

近江牡蛎HSP70基因对溶藻弧菌感染的反应

采用实时荧光定量RT-PCR方法,检测了注射溶藻弧菌(Vibiro alginolyticus)后近江牡蛎鳃,闭壳肌,消化腺,外套膜,心脏以及血细胞中HSP70基因的表达变化。结果显示近江牡蛎这五种器官组织中的HSP70基因表达量均出现显著性高表达,且在鳃、外套膜和血细胞中的HSP70基因表达变化规律表现为典型的时间依赖性。血细胞中,显著高表达的峰值出现在24h,至72h恢复到对照水平,高表达持续时间最长:鳃中表达峰值出现时间较早,在第3h,随后在第12h便恢复到对照水平;外套膜,消化腺以及心脏中的峰值分别出现在6h,6h和3h,而在闭壳肌组织中,没出现显著性高表达。由此可见,近江牡蛎HSP70s可能在机体抗菌免疫过程中起了重要作用。     

Association analysis of HSP70A1A haplotypes with heat tolerance in Chinese Holstein cattle

Single-nucleotide polymorphisms (SNPs) in the coding and untranslated regions of heat shock 70 kDa protein 1A (HSP70A1A), an inducible molecular chaperone that is responsible for cellular protection against heat stress, have been reported as being associated with heat tolerance. A fragment of the HSP70A1A gene was amplified in Chinese Holstein cattle and eight novel mutations were found. We performed comprehensive linkage disequilibrium (LD) and haplotype analyses of the eight SNPs of the HSP70A1A gene and examined their involvement in heat resistance in 600 Chinese Holstein cattle. Our results revealed the presence of significant differences between individuals carrying haplotype 1 and those without haplotype 1 for most of the heat-tolerance traits. Haplotype 1 increased the risk of heat stress; however, association analysis of its combination with haplotype 2 showed the lowest rectal temperature and red blood cell K⁺ level, moderate respiratory rate, and the highest red blood cell NKA level, suggesting a heterozygote advantage in the penetration of the phenotype. Protein expression levels in white blood cells among haplotype combinations further confirmed the hypothesis that heterozygotes for haplotypes 1 and 2 are more sensitive to heat stress. We presume that these mutations may be useful in the future as molecular genetic markers to assist selection for heat tolerance in cattle.     

HSP86 and HSP84 exhibit cellular specificity of expression and co-precipitate with an HSP70 family member in the murine testis

This study extends to the protein level our previous observations, which had established the stage and cellular specificity of expression of hsp86 and hsp84 in the murine testis in the absence of exogenous stress. Immunoblot analysis was used to demonstrate that HSP86 protein was present throughout testicular development and that its levels increased with the appearance of differentiating germ cells. HSP86 was most abundant in the germ cell population and was present at significantly lower levels in the somatic cells. By contrast, the HSP84 protein was detected in the somatic cells of the testis rather than in germ cells. The steady-state levels of HSP86 and HSP84 paralleled the pattern of the expression of their respective mRNAs, suggesting that regulation at the level of translation was not a major mechanism controlling hsp90 gene expression in testicular cells. Immunoprecipitation analysis revealed that a 70-kDa protein coprecipitated with the HSP86/HSP84 proteins in testicular homogenates. This protein was identified as an HSP70 family member by immunoblot analysis, suggesting that HSP70 and HSP90 family members interact in testicular cells. © 1993Wiley-Liss, Inc.     

饥饿对小鼠脑组织中转氨酶、肌酸激酶、乳酸脱氢酶和热休克蛋白70的影响

饥饿处理小鼠1～3天,分别检测脑组织中转氨酶、肌酸激酶、乳酸脱氧酶和HSPT0的变化.结果表明,谷丙转氨酶在饥饿1～3天有逐渐增高的趋势;谷草转氨酶在饥饿1～2天明显升高,但饥饿3天后迅速下降;在正常脑组织中,肌酸激酶的活性较强,饥饿1天后迅速下降,2～3天维持在较低水平;乳酸脱氢酶和HSP70受饥饿影响明显升高.     

Comparative analysis of differential gene expression of HSP40 and HSP70 family isoforms during heat stress and HIV-1 infection in T-cells

Heat shock proteins (HSPs) are a family of cellular proteins involved in a variety of biological functions including chaperone activity. HSPs are classified based on their molecular weight and each family has several isoforms in eukaryotes. HSP40 is the most diverse family acting as a co-chaperone for the highly conserved HSP70 family. Some of the isoforms are reported to be induced during heat stress. Few studies have also highlighted the diverse role of some isoforms in different stress conditions including viral infections. But till date, no study has comprehensively examined the expression profile of different HSP40 and 70 isoforms in either heat stress or HIV-1 infection, a virus that is responsible for the pandemic of AIDS. In the present study, we have compared the mRNA expression profile of HSP40 and HSP70 isoforms during heat stress and HIV-1 infection in a T-cell line and also validated the HIV-1 stress results in peripheral blood mononuclear cells. In case of HSP70, we observed that three isoforms (HSPA1A, HSPA1B, and HSPA6) are highly upregulated during heat stress, but these isoforms were found to be downregulated during the peak of HIV-1 infection. While in case of HSP40, we found that only DNAJA4, DNAJB1, and DNAJB4 showed significant upregulation during heat stress, whereas in HIV-1 infection, majority of the isoforms were induced significantly. Stress-dependent differential expression observed here indicates that different HSP40 and HSP70 isoforms may have specific roles during HIV-1 infection and thus could be important for future studies.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12192-020-01185-y.     

地衣芽孢杆菌对凡纳滨对虾Toll和HSP70基因表达的影响

目的 分析地衣芽孢杆菌对凡纳滨对虾Toll和HSP70基因表达的影响.方法 根据芽孢杆菌的使用量设置测试1组(LDG,用菌量2.0×103 CFU/mL),测试2组(HDG,用菌量1.0 × 104 CFU/mL)及未使用芽孢杆菌的对照组(CG),每7d施菌1次,实验持续24 d.每6d取样对虾肌肉和肝胰腺检测Toll基因和HSP70基因的mRNA相对表达量.结果 对虾肝胰腺Toll基因mRNA表达量在LDG组和HDG组分别较CG组提高了410.05％和78.31％,但在肌肉中CG组的表达量则分别较LDG组和HDG组提高了31.81％和8.43％.可见,芽孢杆菌在一定程度上可提高Toll基因mRNA在对虾肝胰腺的表达,但对其在肌肉中的表达则呈一定的限制作用.在肝胰腺HSP70基因的mRNA表达量,HDG组显著高于CG组和LDG组(P＜0.05),CG组和LDG组的差异无统计学意义(P＞0.05);在肌肉中LDG组和HDG组的HSP70基因mRNA表达分别较CG组提高了26.07％和26.46％,但实验组的组间差异无统计学意义(P＞0.05).结论 在水体中施用芽孢杆菌104 CFU/mL和2.0×103 CFU/mL可分别提高对虾肝胰腺和肌肉HSP70基因的mRNA表达量.     

急性冷应激对牦牛乳腺上皮细胞 HSP70 mRNA 表达的影响

本文主要研究了急性冷应激对牦牛乳腺上皮细胞热休克蛋白７０ （Heat stress protein,ＨＳＰ７０） 表达量的影响。采用实时荧光定量ＰＣＲ 技术,以急性冷刺激１０℃ 为典型研究环境,分析了ＨＳＰ７０ ｍＲＮＡ 表达的变化规律。结果显示,乳腺上皮细胞在１０℃分别冷处理２ ｈ、４ ｈ、６ ｈ 和８ ｈ,其ＨＳＰ７０ ｍＲＮＡ 的表达量变化均不显著（Ｐ ＞０. ０５）;分别在１０℃冷处理２ ｈ、４ ｈ、６ ｈ 和８ ｈ,再复温培养４ ｈ,ＨＳＰ７０ ｍＲＮＡ 的表达量均极显著增加（Ｐ ＜０. ０１）,于６ ｈ 达到峰值;在１０℃先冷处理４ ｈ,然后分别复温２ ｈ、４ ｈ、６ ｈ 和８ ｈ,ＨＳＰ７０ ｍＲＮＡ 的表达量亦均显著增加（Ｐ ＜０. ０１）,并于４ ｈ 达到峰值。结论：急性冷应激诱导牦牛乳腺上皮细胞ＨＳＰ７０ 表达量的增加不是发生在冷处理过程中,而是发生在复温过程中,并且在一定范围内随冷处理时间的增长表达量增高。     

Effect of sericin supplementation on heat shock protein 70 (HSP70) expression,redox status and post thaw semen quality in goat

Cryopreservation results in substantial deterioration of heat shock protein 70 (HSP70) and ultra-structural changes in sperm organelles, resulting in a marked reduction in post-thaw semen quality. The present study was aimed to explicate the effect of sericin supplementation on expression profile of HSP70, redox status and post-thaw semen quality in Barbari goat. Five Barbari bucks were used to collect thirty semen ejaculates by using artificial vagina and each ejaculate was divided into three aliquots to which sericin was supplemented at 0% (Control), 0.25% (T1) and 0.50% (T2). Further, extended semen samples were equilibrated followed by their cryopreservation. Post-thaw semen characteristics, redox status of seminal plasma, enzyme leakage and HSP70 gene/protein expression in spermatozoa were assessed in all the groups. Per cent progressive motile spermatozoa, spermatozoa having intact plasma membrane (HOST + ve) and intact acrosomes in post-thaw spermatozoa were significantly (p < 0.01) higher in T1 and T2 as compared to control. A significant (p < 0.01) reduction in abnormal spermatozoa was found in T1 as compared to T2. Sericin supplementation significantly (p < 0.05) improved the antioxidative status (SOD, GST, CAT), reduced lipid peroxidation (MDA) and also prevented enzyme (ALT, LDH) leakage as compared to control samples. qRT-PCR results revealed that HSP70 mRNA expression was significantly (p < 0.01) upregulated in T1 and T2 group as compared to control. The positive effect of sericin on expression of HSP70 was further confirmed by immunoblotting followed by densitometry revealing higher expression in T1 and T2 compared to control. Inclusion of 0.25% w/v sericin in semen extender ameliorated the post-thaw semen quality by improving antioxidative status and minimizing the leakage of intracellular enzymes. Sericin supplementation had a beneficial effect on HSP70/HSP70 mRNA expression either by induction or by protection of HSP70/HSP70 mRNA as evident from the gene expression and immunoblotting studies.     

热应激对小鼠器官指数、小肠损伤及胃HSP70 mRNA表达的影响

目的探讨热应激对小鼠器官指数、小肠形态、胃黏膜HSP70 mRNA表达量及糖代谢相关激素的影响。方法采用单因子实验设计,将年龄和体重相近的18只KM小鼠随机分为对照组和热应激组,分别测定心、肝、脾、肺、肾重量,小鼠胃黏膜HSP70 mRNA表达量、血浆中胰岛素和胰高血糖素浓度以及十二指肠和空肠的绒毛高度、隐窝深度,并对肝脏、十二指肠和空肠进行病理组织学检查。结果与结论热应激对小鼠器官指数无影响,可显著提高小鼠胃黏膜HSP70 mRNA表达量,降低血浆中胰岛素的含量,并造成小鼠肝脏、十二指肠和空肠严重损伤。     

Possible involvement of HSP70 in pancreatic cancer cell proliferation after heat exposure and impact on RFA postoperative patient prognosis

PurposeAs an alleviative treatment measured in patients with unresectable advanced pancreatic cancer, radiofrequency ablation (RFA) needed more clinical data to prove its advantages and to explore limitations in its utilization. This study was determined to observe the efficacy of RFA, and to explore its impact on perioperative periphery carcinoma as well as the normal pancreatic tissues.MethodsClinical data of 32 patients with pancreatic cancer accepted RFA surgery were collected. Followed up patients’ pain degree and the changes in serum tumor markers CA19-9 and CA 242 before and after surgery. Ex vivo, gave human pancreatic cancer cell line PANC-1 heat treatment to simulate the heat exposure condition periphery carcinoma was experienced during RFA surgery, and to observe the proliferation rate and HSP70 expression change compared with control group.ResultsOf the 32 patients, 1 died of upper gastrointestinal hemorrhage, and 29 survived for more than 5 months, 2 of which for more than 16 months. The average CA19-9 and CA 242 levels of the patients were significantly decreased in 3 months after surgery (t = 9.873, 5.978, P < 0.001). During in vitro experiments, the proliferation rate of PANC-1 cells after heating was significantly increased, accompanied with the increased HSP70 expression. The addition of HSP70 inhibitor can inhibit the rise of proliferation after heat therapy.ConclusionUtilizing RFA treat patients with unresectable advanced pancreatic cancer, could effectively relieve the pain, decline jaundice, and deduce tumor marker levels significantly. However, it failed to extend the long-term survival rate of the patients significantly. This study found that a higher proliferative rate accompanied with a higher HSP70 expression level were observed on in vitro cultured pancreatic carcinoma cells after heat treatment, which could be altered by HSP70 inhibitor. And these findings indicated that the heat exposure might impact periphery carcinoma during RFA surgery and HSP70 might play an important role in patients’ prognosis.