猪肌肉生长抑制素基因5′调控区T→A突变与生长性状的关系分析

生长速度是猪育种中选择的主要目标性状之一,鉴定影响生长速度的QTL或基因具有重要的实践意义和理论价值.对猪肌肉生长抑制素基因(myostatin,MSTN)5′调控区TA突变与生长性状的关系进行了分析.突变所产生的基因型的判定采用PCR-RFLP方法进行.猪生长性状的测定指标包括60日龄体重(body weight at 60d,BW60)、前期平均日增重(average daily gain of stage one,ADG1)、后期平均日增重(average daily gain of stage two,ADG2)和全期平均日增重(average daily gain of whole stage,ADG),所采用的记录数分别来自165,276,275和275头无亲缘关系个体.采用SAS软件包的GLM程序对MSTN基因型与猪生长性状的关系进行分析.结果表明,MSTN基因型对猪BW60,ADG1和ADG2的效应不显著(P＞0.1),对ADG2的效应在剔除基因型与品种的互作效应以后几乎达到显著水平(P＝0.0522),突变等位基因的携带者(基因型为TA)具有较高的后期平均日增重.     

Polymorphisms in the 5′ regulatory region of myostatin gene are associated with early growth traits in Yorkshire pigs

Myostatin is a negative regulator of skeletal muscle mass. The present study cloned the 5' regulatory region of porcine myostatin gene, screened its polymorphisms and analyzed their associations with early growth traits in Yorkshire pigs. The results indicated that a fragment length polymorphism and a polymorphism concerning two nucleotide changes exist in the 5' regulatory region of porcine myostatin gene. At sites 435 and 447, allele A and allele B have the haplotypes of A-G and G-A, respectively. The allelic frequency of B is 0.475 in Yorkshire pigs. No homozygous BB genotype was detected in 9 Laiwu Black pigs. Allele B was found to have positive effect on body weight on day 21 (BW21) (P＜0.01), body weight on day 28 (BW28) (P＜0.05), body weight on day 70 (BW70) (P＜0.05), average daily gain from birth to 21 d (ADG1) (P＜0.01), average daily gain from birth to 28 d (ADG2) (P＜0.05) and average daily gain from 21 d to 70 d (ADG3) (P＜0.01), respectively. The additive effect of allele B on BW21, BW28, BW70, ADG1, ADG2 and ADG3 was 0.596±0.205 kg (P=0.0041), 0.498±0.200 kg (P=0.0136), 1.409±0.551 kg (P=0.0112), 28.39±9.74 g P=0.0041), 17.78±7.15 g (P=0.0136) and 37.00±16.92 g (P=0.0304), respectively, whereas its effect on average daily gain from 28 d to 70 d (ADG4) was not significant (P＞0.1), although BB individuals are superior in average daily gain to AA and AB.     

Polymorphisms in the 5 ′ regulatory region of my-ostatin gene are associated with early growth traits in Yorkshire pigs

Myostatin is a negative regulator of skeletal muscle mass. The present study cloned the 5′ regulatory region of porcine myostatin gene, screened its polymorphisms and analyzed their associations with early growth traits in Yorkshire pigs. The results indicated that a fragment length polymorphism and a polymorphism concerning two nucleotide changes exist in the 5′ regulatory region of porcine my-ostatin gene. At sites 435 and 447, allele A and allele B have the haplotypes of A-G and G-A, respec-tively. The allelic frequency of B is 0.475 in Yorkshire pigs. No homozygous BB genotype was detected in 9 Laiwu Black pigs. Allele B was found to have positive effect on body weight on day 21 (BW21) (P<0.01), body weight on day 28 (BW28) (P<0.05), body weight on day 70 (BW70) (P<0.05), average daily gain from birth to 21 d (ADG1) (P<0.01), average daily gain from birth to 28 d (ADG2) (P<0.05) and av-erage daily gain from 21 d to 70 d (ADG3) (P<0.01), respectively. The additive effect of allele B on BW21, BW28, BW70, ADG1, ADG2 and ADG3 was 0.596±0.205 kg (P=0.0041), 0.498±0.200 kg (P=0.0136), 1.409±0.551 kg (P=0.0112), 28.39±9.74 g P=0.0041), 17.78±7.15 g (P=0.0136) and 37.00±16.92 g (P=0.0304), respectively, whereas its effect on average daily gain from 28 d to 70 d (ADG4) was not significant (P>0.1), although BB individuals are superior in average daily gain to AA and AB.     

锌指核酸酶技术制备肌肉生长抑制素基因敲除的五指山小型猪成纤维细胞

敲除猪肌肉生长抑制素(Myostatin, MSTN)基因可能提高猪瘦肉率, MSTN基因敲除猪也可作为相关疾病的动物模型。文章利用锌指核酸酶(Zinc-finger nucleases, ZFNs)技术敲除五指山小型猪胎儿成纤维细胞MSTN基因, 为制备MSTN基因敲除猪奠定基础。ZFNs质粒或编码ZFNs的mRNA均能高效敲除MSTN基因, 使用ZFNs mRNA能直接得到MSTN+/-和MSTN-/-两种基因型的细胞克隆。DNA序列测定与分析发现, 细胞克隆的突变类型多为ZFNs作用靶位点处不大于10 bp的碱基插入或缺失(92.18 %); 氨基酸预测发现, 突变型MSTN基因的终止密码子常常提前出现。将MSTN基因敲除的细胞进行体细胞核移植(Somatic cell nuclear transfer, SCNT)发现, 胚胎体外早期发育潜力与野生型无显著差异, 表明这些细胞可用于后续MSTN基因敲除猪的制备。     

黄颡鱼MSTN基因多态性及其与生长性状的相关性分析

肌肉生长抑制素基因(Myostatin,MSTN)属于转化生长因子β家族,其主要功能是负向调控肌肉的生长发育。采用PCR-SSCP技术对黄颡鱼MSTN基因进行单核苷酸多态性检测和分型,并与其生长性状进行关联分析。结果表明,在第一内含子部分检测到1个缺失位点和2个突变位点(T1003del、G1022A和T1063G),基因型分别为:AA、AB、CC、CD和DD;在第三外显子部分检测到1个突变位点(T132C),基因型分别为:EE和EF。关联性分析表明,AA基因型个体的全长、体长、体高、体厚、头长和体重显著大于CD和DD基因型(P<0.05),AA基因型雌性个体的全长、体长、体高、体厚、头长、尾柄高、尾柄厚和体重也显著大于DD基因型(P<0.05)。由此推断,AA基因型是影响雌性黄颡鱼生长性状的有利基因型,DD基因型是影响雌性黄颡鱼生长性状的不利基因型,可以尝试利用这两个位点对雌性黄颡鱼进行标记辅助选育。     

应用VCE4.0估计长白猪生长性状的遗传参数

应用PEST和VCE4.0对温氏长白猪的主要生长性状进行了遗传分析。利用温氏水台原种猪场1998～2002年的6344头长白猪生长性能测定记录,运用多性状动物模型REML方法估计主要生长性状的遗传参数。估计遗传力的变化范围为0.207～0.493,性状达30kg体重日龄（AGE30）、达100kg体重日龄（AGE100）、30～100kg平均日增重（ADG）和100kg体重活体背膘厚（FAT）的估计遗传力分别是0.207、0.396、0.304和0.493;FAT/ADG、FAT/AGE100、ADG/AGE100、ADG/AGE30和AGE30/AGE100的各自遗传相关为-0.343、0.180、-0.941、-0.48和0.745;它们的表型相关分别为-0.139、0.138、-0.829、-0.026 和 0.565;AGE30、AGE100、ADG和FAT的窝效应估计结果分别为0.194、0.156、0.157和0.043。     

山东部分地方猪种MyoG基因5''''端遗传多态性及遗传效应分析

采用PCR-SSCP方法对莱芜猪、大蒲莲猪、沂蒙黑猪和里岔黑猪的肌细胞生成素(myogenin,MyoG)基因5'端进行单核苷酸多态性检测,并分析MyoG基因对猪的初生重、60日龄重、210日龄重,60～210日龄平均日增重和背膘厚的影响。根据猪MyoG基因5'端的DNA序列(U14331)设计3对引物,发现5-2引物对扩增的片段有多态性,且存在3种基因型(AA、AB、BB),并对纯合子进行测序,发现2080位G→A突变。χ2独立性检验表明,基因型频率在莱芜猪和大蒲莲猪与沂蒙黑猪和里岔黑猪间存在显著差异(P<0.05)。固定效应模型分析结果表明,初生重基因型间差异显著(P<0.05),而60日龄重、210日龄重、60～210日龄平均日增重和背膘厚基因型间差异不显著(P>0.05)。最小二乘分析结果表明,BB基因型与其他两种基因型比较有较小的初生重和日增重,同AA和AB基因型比较差异极显著(P<0.01),3种基因型在初生重和60～210日龄日增重的大小排列顺序一致,为AA>AB>BB。因此,推测基因型对个体的初生重和日增重存在一定的影响,选择带有A等位基因的个体有望提高个体的初生重和日增重。     

猪个体基因杂合度对生长性状的影响

逐个测定 139头F2 代杂交猪 (Susscrofa) (英系大白猪×梅山猪 )的初生重、180日龄活重 (LWT180 )、日增重(ADG)、相对生长率 (RGR)和KR值等 5个生长性状 ,对每一个体分别进行 2 4个微卫星座位的基因分型。对性别、家系和标记与生长性状的关系进行分析 ,结果显示 :它们与这些生长性状没有显著关联 (P >0 0 5 )。以 2 4个中性微卫星座位的杂合度来表示个体基因杂合度 ,比较猪生长性状在各个体基因杂合度水平间的差异 ,并分析其变化规律。结果表明 :初生重从个体基因杂合度水平为 1( 1 5 185kg)时至杂合度水平为 2 ( 1 30 6 3kg)时达性状最低值 ,之后随基因杂合度水平的增加而增加 ;180日龄活重、日增重、相对生长率和KR值等 4个重要经济性状随个体基因杂合度的增加呈近似于正弦函数曲线变化 ,在个体基因杂合度水平 2 (杂合度为 0 5 796 )时的性状值最高 ,之后随着个体基因杂合度的增加而降低。同一性状在不同的个体基因杂合度水平表型值不同 ,在某一特定的杂合度水平时达最高 ,提示在猪选配时将个体基因杂合度控制在恰当的水平 ,可以使目的性状的表型值最高。     

编辑MSTN半胱氨酸节基元促进两广小花猪肌肉生长

肌生长抑制素(myostatin,MSTN)是转化生长因子β(transforming growth factor-β,TGF-β)家族成员之一,是一种肌肉生长抑制因子。解除MSTN的生长抑制功能是提高畜禽肌肉产量的一种有效途径。TGF-β的半胱氨酸节结构基元(cystine knot motif)能够稳定MSTN蛋白结构,对MSTN生物学功能的发挥具有重要调控作用。本研究应用CRISRP/Cas9基因编辑技术在两广小花猪肾细胞(Liang Guang small spotted pig kidney cells,LPKCs)中对MSTN基因外显子3进行编辑,破坏了其半胱氨酸节基元,以解除MSTN对靶基因的抑制功能。将流式分选获得的混合阳性MSTN编辑LPKCs作为供体细胞进行核移植和胚胎移植,获得8头MSTN基因编辑两广小花猪仔猪,其中2头存活至10日龄,经鉴定这2头均为基因编辑杂合子,它们在构成MSNT蛋白半胱氨酸节基元的两个半胱氨残基C106和C108编码序列附近分别发生碱基的缺失与替换,导致移码突变,使C106和C108突变为其他氨基酸。MSTN基因编辑两广小花猪杂合子肩部和臀部肌肉较为发达。H&E切片分析显示,MSTN基因编辑猪肌纤维横截面积显著减少,肌纤维数量显著增多。Western Blot分析结果显示,C106和C108缺失对MSTN蛋白表达无显著性影响,但显著促进其靶基因Myf5、MyoD和Myogenin等成肌相关因子的表达。本研究获得的基因编辑猪模型没有造成MSTN表达完全缺失,可保留MSTN其他生物学功能,在促进两广小花猪肌肉生长的同时还消除了MSTN完全缺失可能对小花猪造成的潜在影响。     

牛肌肉生长抑制素基因突变的遗传效应与育种应用

肌肉生长抑制素（myostatin,MSTN）基因主要在骨骼肌中表达,参与调控骨骼肌的生长发育。MSTN基因在不同物种中具有极强的进化保守性,同时还具有较多的突变多态性。在牛的不同品种中,存在不同位点的有义突变,突变型牛均表现为骨骼肌发达,呈现双肌表型,生长速度与产肉率显著提高。同时,该基因突变也引起显著的生理性遗传效应。对国内外肉牛的MSTN基因突变类型、突变后遗传效应及在肉牛育种应用等方面作了重点阐述,以期为我国地方品种肉牛改良和选育研究提供参考。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.