狼山鸡保种群不同世代MHC BF基因遗传多样性研究

通过Illumina平台Miseq重测序方法分析不同世代(G0、G5、G10、G12、G14、G15、G17)的MHC BF1和BF2基因座位多态性,揭示狼山鸡不同世代间MHC BF基因遗传多样性,为更好地监测保种效果提供科学的决策依据。在狼山鸡7个世代中分别鉴定出MHC BF1、BF2基因59和34个SNPs。狼山鸡MHC BF1基因观察杂合度(Ho)、期望杂合度(He)和多态信息含量(PIC)在G10、G12、G14保持稳定,Ho在不同世代变化情况为G17G15G14、G12、G10G5G0,He和PIC在不同世代变化情况为G17、G15G14、G12、G10G5G0,MHC BF2基因的Ho在G10、G14、G15保持稳定,其他世代变化情况为G17、G12G15、G14、G10G0、G5;He和PIC在G12、G14、G15和G17保持稳定,在其他世代变化情况为G17、G15、G14、G12G10G5、G0。从本研究结果看,狼山鸡不同世代MHC BF1和BF2遗传多样性变化规律不太一致,但总体规律符合波动选择(时空变换的选择)假说,随着时间的推移MHC BF基因遗传多样性变得更为丰富,在某些连续几个世代群体遗传多样性基本保持稳定,这可能与MHC BF基因功能有关。     

畜禽主要组织相容性复合体与抗病性的研究

随着分子生物学技术的发展,人们越来越重视育种的效率,对主要组织相容性复合体研究的进一步深入,阐明了它在免疫应答、多种疾病的预防方面起到了一定的作用,主要介绍了主要组织相容性复合体的概念以及它与抗病性的关系,最后对主要组织形容性复合体的发展进行了展望。     

长江草鱼群体MHC Class ⅡB的多态性和进化分析

克隆及测序草鱼(Ctenopharyngodon idella)长江3个群体的主要组织相容性复合体(MHC)Class II B基因编码β1和β2区的第2和第3个外显子及两个外显子之间的内含子,分析了草鱼MHC的进化模式和种群遗传结构。结果显示:实验共定义了34个等位基因,每条序列包括长为130~136 bp的第2个外显子,长为218 bp的第3个外显子以及长81~371 bp的内含子。序列分析揭示,第2个外显子有106个核苷酸变异位点(78%)和40个氨基酸变异位点(88%),而第3个外显子有100个核苷酸变异位点(45%)和41个氨基酸变异位点(56%),β1变异要大于β2区。用β1和β2区序列分别构建的邻接(NJ)系统树均显示5个具有高支持率的谱系,结合序列变异特点和内含子长度,推测草鱼至少存在5个MHC Class II座位。分别计算β1的肽结合位点(PBR)、非肽结合位点及β2的非同义替换率(dN)和同义替换率(dS),PBR的dN/dS为2.03(P<0.05),非肽结合位点和β2则小于1,表明草鱼MHC受到歧化选择作用。根据等位基因在群体中的分布频率作分子方差分析(AMOVA),得出FST为0.37%,提示长江草鱼MHC没有遗传分化。     

星斑川鲽MHCⅡ恒定链Ii基因的克隆和表达特性

为了研究星斑川鲽MHCⅡ类分子的作用及调控机制,实验通过SMART-RACE技术克隆得到了星斑川鲽MHCⅡ恒定链(MHCⅡIi)的全长cDNA序列,其长度为1766 bp,包含135 bp的5′非编码区、837 bp的开放阅读框和794 bp的3′非编码区。该基因共编码279个氨基酸。理论分子量为30.848 ku,等电点为6.89。与已知物种MHCⅡIi进行同源性比对,结果与狼鲈、紫红笛鲷和鳜关系较近,同源性均为79%。利用quantitative realtime PCR(qRT-PCR)技术检测了MHCⅡIi在星斑川鲽不同组织中的表达,以及爱德华氏菌感染前后对该基因在不同组织中表达水平的影响,结果显示:在脾脏、头肾、肝脏、后肠、性腺、心脏、血液、鳃和肌肉组织中,MHCⅡIi mRNA均有表达,但在表达量上有明显差异,脾脏和头肾组织相对表达水平较高,鳃、血液、肌肉、心脏和性腺中的表达水平较低。病原感染后,免疫相关组织脾脏和头肾的表达水平升高最明显,肝脏和后肠的表达水平也略有升高,但变化不明显。本研究可为星斑川鲽MHCⅡ类分子的作用机理提供理论依据,同时为海水养殖鱼类的抗病遗传育种工作提供研究基础。     

中国泰和乌骨鸡MHC与其繁殖性能相关的研究

实验采用四种限制性内切酶（ＨｈａＩ、ＥｃｏＲＶ、ＨａｅⅢ、Ｘｂａｉ）,运用ＰＣＲ－ＲＦＬＰＳ的方法,研究了泰和乌骨鸡ＭＨＣ分子遗传多态笥,探讨了各基因型与繁殖性能的相关关系,结果表明：四种酶切区别的Ｂ－ＬＩＩＢβ１外元的多态笥及ＭＨＣ基因组合型对蛋重及受精蛋孵化率影响差异均不显著。ＥｃｏＲＶ及ＸｂａＩ酶切多态性对产蛋数的多态性及ＭＨＣ基因组合型对蛋重及受精蛋孵化率影响差异均不显著。ＥｃｏＲＶ及Ｘｂ     

家禽MHC的结构与遗传效应

MHC是重要的免疫分子,能够稳定的遗传,并且与疾病有明显的相关性。研究家禽MHC分子特性是阐明机体免疫机制的重要途径之一。本文就家禽MHC基因结构与多态性、抗原递呈作用以及其与疾病和生产性能的相关性等方面进行了阐述。     

Expression of MHC Ⅱ on rat corneal keratocytes is inhibited by special siRNA targeting CⅡTA

AIM: To investigate the feasibility to inhibit the expression of MHCⅡ by special siRNA targeting class Ⅱ major histocompatibility complex (MHC Ⅱ) transactivator (CⅡTA), which might regulate MHC Ⅱexpression for suppressing immune rejection. METHODS: Five different siRNA were designed, synthesized and transfected into freshly isolated rat corneal keratocytes. At 24 hours posttransfection, the changes of MHC Ⅱexpression were detected by flowcytometry, and the mRNA abundance of CⅡTA and MHC Ⅱ was measured by FQ-PCR after inducing with recombinant rat interferon-gamma (IFN-γ). RESULTS: Different siRNA showed different reduction in MHC Ⅱ and CⅡTA expression compared with the control (P<0.01). Among the five groups, the siRNA-4 was the most efficient. The mRNA content of CⅡTA and MHC Ⅱ were reduced by 95.10%±1.25% and 82.70%±1.95% respectively and the expression of MHC Ⅱ was inhibited by over 80% in siRNA-4 group at 24 hours posttransfection. CONCLUSIONS: The special siRNA targeting to CⅡTA inhibits CⅡTA mRNA and further inhibits its regulation of MHC Ⅱ molecular expression. The blockade of MHC Ⅱ by siRNA may be useful for further studying allogeneic corneal limbal transplantation.     

绵羊MHCⅡ类基因遗传多态性研究进展

主要组织相容性复合体(major histocompatibility complex,MHC)是广泛存在于脊椎动物体内的一类高度紧密连锁的基因群,绵羊MHC又称为绵羊淋巴细胞表面抗原(ovine lymphocyte antigen,OLA),位于绵羊20号染色体上,分为Ⅰ类、Ⅱ类和Ⅲ类区域,其中MHCⅡ类基因具有高度的基因多态性.文章综述了绵羊MHCⅡ类基因的分子结构及遗传多态性,重点总结了近十年来国内外不同绵羊品种MHCⅡ类基因遗传多态性的研究进展,并对绵羊MHCⅡ类基因未来的研究重点进行了展望.     

Comparative Analysis of mRNA Expression of Surface Antigens between Histiocytic and Nonhistiocytic Sarcoma in Dogs

Background

Definitive diagnosis of histiocytic sarcoma (HS) in dogs is relatively difficult by conventional histopathological examination because objective features of HS are not well defined.

Hypothesis

Quantitative analysis of mRNA expression of selected cellular surface antigens (SAs) specific to HS in dogs can facilitate objective and rapid diagnosis.

Animals

Dogs with HS (n = 30) and dogs without HS (n = 36), including those with other forms of lymphoma (n = 4), inflammatory diseases (n = 6), and other malignant neoplasias (n = 26).

Methods

Retrospective clinical observational study. Specimens were collected by excisional biopsy, needle core biopsy, or fine needle aspiration. To determine HS detection efficacy, mRNA expression levels of selected SAs specific to HS in dogs, including MHC class IIα, CD11b, CD11c, and CD86, were quantitatively analyzed using real‐time quantitative polymerase chain reaction.

Results

Each SA mRNA expression level was significantly higher in HS dogs than in non‐HS dogs (P = .0082). Cutoff values for discriminating between HS and non‐HS dogs based on these expression levels were calculated on the basis of receiver‐operating characteristic analysis. Accuracy of the cutoff values, including MHC class IIα, CD11b, CD11c, and CD86, was 87.9, 86.4, 86.4, and 84.8%, respectively.

Conclusions and Clinical Importance

Our results suggest that quantitative analysis of mRNA expression of the selected SAs could be an adjunctive diagnostic technique with high diagnostic accuracy for HS in dogs. Substantial investigation is required for exclusion of diseases with similar cell types of origin to lymphoma.     

Cloning and Sequence Analysis of Major Histocompatibility Complex Ⅰ Gene from Goose F1 of Fast-growth Lines

According to the multiple alignments identified major histocompatibility complex Ⅰ (MHC Ⅰ) gene conserved sequence registered in GenBank from the family ducks (Anatidae) anser waterfowl, a pairs of specific primers for the fragments of MHCⅠgene of goose F1 from fast-growth lines were designed and synthesized by Primer Premier 5.0. Using the genome DNA of goose F1 from fast-growth lines, the target gene fragment was obtained by PCR. To conduct sequencing of the fragments of MHCⅠgene of goose F1 from fast-growth lines and make sequence alignment and analysis of protein structure and function by bioinformatics, and research the characteristics of MHCⅠgene of goose F1 and the physicochemical properties of the protein. Bioinformatics was analyzed the nucleic acid data, deduced amino acid sequence and phylogenetic trees. The result of sequence analysis showed that the fragments of MHCⅠgene of goose F1 from fast-growth lines was 1036 bp in length, which coded 96 amino acids polyprotein. The homology were 93% and 83% with MHC Ⅰ gene and coding sequence of Wulong goose in NCBI respectively. There were 72 different bases sequence and 16 amino acids change. There also was higher homology with other poultry, and existed genetic relationship of Siji goose > chickens > ducks.The homology segment sequences corresponding to the fragments of MHCⅠ gene of goose F1 coded 96 amino acids protein, which molecular weight, PI, positively or negatively charged amino acid, estimated half-life, instability index, aliphatic index and average hydrophobicity were 11.342 ku, 5.32, 14, 17, 2.8 h, 34.92, 42.81, -1.066, respectively, and appeared 9 B cell epitopes, but contained no signal peptide. These results indicated that the protein for hydrophilic non-secreted proteins, had the high immunogenicity. In addition, The protein structure study indicated that alpha-helix, beta-sheet, beta-turn and random coil were 31.25%,16.67%, 14.58% and 37.50%, respectively. There existed amino terminal domain and carboxyl terminal domain in the tertiary structure. Therefore, MHC gene had significant difference between species and populations of individuals by the pathogen pressure in environment, and there were the interaction between polymorphism of MHC molecules and the diversity of antigenic peptide. MHC determined the differences of individual susceptibility to disease, and could be treated as a candidate gene for disease resistance.