Rice Storage Proteins:Focus on Composition,Distribution,Genetic Improvement and Effects on Rice Quality

Rice storage proteins(RSPs)are plant proteins with high nutritional quality.As the second largest type of storage substance in rice,it is the main source of protein intake for people who consume rice as a staple food.The content and type of RSPs affect the appearance,processing quality and eating quality of rice.These effects involve the distribution of RSPs in rice grains as well as the interactions of RSPs with other components such as starch in rice grains.In the past two decades,some progress has been made in the genetic improvement of RSPs.However,the determination mechanism of protein content and composition in rice is still unclear,and the mechanism of the effect of RSPs on rice quality has not been elucidated.In this review,the composition,biosynthesis and distribution of RSPs,and quantitative trait loci mapping and cloning of RSP genes are summarized,the research progress of the influence of RSPs and their components on rice quality are reviewed,and the research directions in the future are proposed.

Considering calcium-binding proteins in invertebrates: multi-functional proteins that shape neuronal growth

Calcium is a critical second messenger molecule in all cells and is vital in neurons for synaptic transmission.Given this importance,calcium ions are tightly controlled by a host of molecular players including ion channels,sensors,and buffering proteins.Calcium can act directly by binding to signaling molecules or calcium’s effects can be indirect,for example by altering nuclear histones.

Genetic Diversity of Seed Storage Proteins in Different Ecotype Varieties of japonica Rice and Its Application

One hundred and fifteen varieties （including cultivars and lines） with different ecotypes in japonica rice （Oryza sativa L.） were analyzed for endosperm storage proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS-PAGE） to estimate their genetic diversity for the purpose of genetic improving and variety identification. Nineteen types of profile were identified according to 1） presence/absence of 65 kDa bands, 2） staining intensity of 70, 60, 57, 37-39, 22-23, 13 and 10 kDa bands, 3） migration velocity of 35 kDa （α-4） and 4） band number at 57 kDa location. An unweighted-pair group average method with arithmetic mean （UPGMA） dendregram based on the cluster analysis of genetic similarity of the protein bands showed a small genetic variation among the tested materials, with the similarity coefficients varying between 0.75 and 1.00. Three distinct groups were identified from the cluster analysis of the rice varieties studied at the similarity coefficient level of 0.894. The first group included eight varieties with high amylose content, the second group contained fifteen varieties with high protein content, and the third group had the remaining ninety-two varieties, which accounted for 80% of the total materials. Clear relationship between ecotypes distinguished by maturity and groups revealed by cluster analysis was not found in this study. Only the group of high amylose linked with medium-maturity medium japonica ecotype. The bands of 70 kDa and 65 kDa can be used as protein markers to identify F1 seed purity of japonica hybrid rice Liuyanyou 422.

Analysis of Genetic Variation of Seed Proteins in the Genus Vigna and among Its Relatives Cultivated in China

The genetic variation of seed proteins was assayed by SDSPAGE for 24 cultivars belonging to 5 species in Vigna and 7 species in its 7 relative genera cultivated in China. There were 48 polymorphic subunit bands discriminated from electrophoretic profiles. Two dendrograms were constructed by UPGMA cluster analyses using PHYLIP3.6 respectively. Variation among genera or species was larger than that among lower taxonomic categories level. Little variation among cuhivars of yardlong bean （Vigna sesquipedalis ） and small variation of lablab （ Lablab purpureus）, pea （Pisum sativum）, or sword bean （Canavalia gladiata）, but large variation of soybean or rice bean in their origin of China were all revealed. The seed proteins profiles of traditionally regarded as typical species in Vigna such as yardlong bean, rice bean and small bean were more similar than mungbean （Vigna radiata） and black gram （Vigna mungo） were. Mungbean and black gram had distinct seed proteins pattern, they should be of two species.

Genetic variation of circHIBADH enhances prostate cancer risk through regulating HNRNPA1-related RNA splicing

The current study aimed to investigate associations of circRNAs and related genetic variants with the risk of prostate cancer(PCa)as well as to elucidate biological mechanisms underlying the associations.We first compared expression levels of circRNAs between 25 paired PCa and adjacent normal tissues to identify riskassociated circRNAs by using the MiOncoCirc database.We then used logistic regression models to evaluate associations between genetic variants in candidate circRNAs and PCa risk among 4662 prostate cancer patients and 3114 healthy controls,and identified circHIBADH rs11973492 T>C as a significant risk-associated variant(odds ratio=1.20,95%confidence interval:1.08-1.34,P=7.06×10^(-4))in a dominant genetic model,which altered the secondary structure of the corresponding RNA chain.In the in silico analysis,we found that circHIBADH sponged and silenced 21 RNA-binding proteins(RBPs)enriched in the RNA splicing pathway,among which HNRNPA1 was identified and validated as a hub RBP using an external RNA-sequencing data as well as the in-house(four tissue samples)and publicly available single-cell transcriptomes.Additionally,we demonstrated that HNRNPA1 influenced hallmarks including MYC target,DNA repair,and E2F target signaling pathways,thereby promoting carcinogenesis.In conclusion,genetic variants in circHIBADH may act as sponges and inhibitors of RNA splicing-associated RBPs including HNRNPA1,playing an oncogenic role in PCa.

Prokaryotical expression of structural and non-structural proteins of hepatitis G virus

AIM To study the epitope distribution of hepatitis G virus (HGV) and to seek for the potential recombinant antigens for the development of HGV diagnositic reagents.METHODS Fourteen clones encompassing HGV gene fragments from core to NS3 and NS5 were constructed using prokaryotic expression vector pRSET and (or)pGEX. and expressed in E. coli. Western blotting and ELISA were used to detect the immunoreactivity of these recombinant proteins.``RESULTS One clone with HGV fragment from core to El(Gl). one from E2 (G31), three from NS3 (G6, G61, G7),one from NS5B (G821) and one chimeric fragment from NS3and NS5B (G61 821) could be expressed well and showed obvious immunoreactivity by Western blotting.One clone with I-KGV framment from NS5B (G82) was also well expressed, but could not show immunoreactivity by Western blotting. No obvious expression was found in the other six clones. All the expressed recombinant proteins were in inclusion body form, except the protein G61 which could be expressed in soluble form. Further purified recombinant proteins Gl, G,31, G61, G821 and G61 821were detected in indirected ELISA as coating antigen respectively. Only recombinant Gl could still show immunoreactivity, and the other four recombinant proteins failed to react to the HGV antibody positive sera.Western blotting results indicated that the immunoactivity of these four recombinant proteins were lost during purification.``CONCLUSION Core to El, E2. NS3 and NS5 fragment of HGV contain antigenic epitopes, which could be produced in prokaryotically expressed recombinant proteins. A high. yield recombinant protein (Gl) located in HGV core to E1 could remain its epitope after purification, which showed the potential that G1 could be used as a coating antigen to develop an ELISA kit for HGV specific antibody diagnosis.

Hepatocellular transport proteins and their role in liver disease

MOLECULAR PHYSIOLLGY OF HEPATOCELLULAR TRANSPORT PROTEINS Basolaferal transport systems Na+-dependent bile salt uptake Uptake of bile salts into the liver was first isolated perfused rat liver[1],isolated hepatocyte cultures and basolateral plasma membrane vesicles [2,4].

Genetic Polymorphism of Milk Protein and Their Relationships with Milking Performances in Chinese Yak

The milk protein polymorphisms were typed by polyacrylamide gel electrophoresis (PAGE)from 109 Maiwa and 100 Jiulong yaks, and the relationships among milk protein polymorphisms,milking traits and milk protein compositions were studied. The results showed thatβ-CN,κ-CN andα-La were monomorphic,αs1-CN andβ-Lg were polymorphic, the dominantgenes were αs1-CN D and β-Lg E,respectively. The frequencies of αs1-CN D were 0.8073and 0.6000 and β-Lg E were 0.9770 and 0.9700 in two populations respectively.The meanheterozygosities were 0.1021 and 0.1867 in two populations. No significant effects onmilking traits and milk protein compositions were observed except for αs1-CN locus onfat percentage in Jiulong yak.

Genetic Analysis of the P1 Region of Human Enterovirus 71 Strains and Expression of the 55 F StrainVP1 Protein

Enterovirus 71 (EV71) is a member of the Entero-virus genus of the Picornaviridae family and is the major cause of Hand, foot, and mouth disease (HFMD) in children. Different strains from Gansu were cloned and the P1 protein was sequenced and analysed. Results indicate that there are three kinds of EV71 infections prevalent in Gansu. The VP1 protein from one of these strains, 55F, was expressed. The recombinant protein was expressed with high level and reacted specifically with the EV71 patient antibody, the recombinant protein was also applied to raise antiserum in rabbits and after the fourth injection a high titer of antiserum was detected by ELISA assay. These data are useful for further clarification of prevalent EV71 strains in the north of China at the molecular level and provide a basis for EV71 diagnosis.

Efficient and Soluble Expression of α Protein of Clostridium perfringens and Preparation of Genetic Engineering Subunit Vaccine

[Objective] The paper was to develop genetic engineering vaccine that can express α exotoxin antigen protein efficiently without destroying its immunogenicity for preventing and controlling the diseases caused by Clostridium perfringens. [Method] Efficiently expressed soluble recombinant α protein was obtained from Escherichia coli expression system by optimizing codon,removing signal peptide,selecting sequences with better hydrophilicity and antigenicity,and optimizing expression conditions. [Result] Mice obtained higher serum antibody level when immunized by α protein,and the immune protection rates against type A,type B,type C and type D C. perfringens were 100%,90%,85% and 90%,respectively. The antibody titer of mice within 7-14 d after the third immunization reached the peak. [Conclusion]The α protein has good immunogenicity,and can be further used to develop genetic engineering subunit vaccines for preventing C. perfringens.

COVID-2019 Genome Sequence Analysis: Phylogenetic Molecular Evolution and Docking of Structural Modelling of Receptor Binding Domain of S Protein in Active Site of ACE2

Meanwhile the outbreak of the Covid-19 since December, 2019 in China, it has killed more than a hundred thousand of people of all ages and sex across the globe in a short span of time. On the bases of this study the nearest family member of the virus and its receptor binding domain of S protein including its model structure and function of its active sites were naked through Multiple Sequence Alignment, modelling and molecular docking software accordingly its repository genome databases. The virus was genetically associated and molecular evolutionary related with (RaTG13) and it scores 96.12% homology with 99% query coverage followed by bat-SL-CoVZC45 and bat-SL-CoVZXC21 notch 89.12% and 88.65% respectively. However, SARS and MERS corona type virus those outbreak earlier respectively less likely family members of 2019-nCoV. Though the virus has a close genetic association with those previous SARS coronaviruses, and certainly the spike protein used as a binding receptor to fight against human receptor protein of ACE 2, but on the basis of FRODOC and HDOCK server analysis multi favorable active sites of S protein was discovered such GLN493 shown as a finest key in both model and possessed a unique traits on it resulting unexpected rate of transmission and number of people died while compared to the previous one. TYR500, ASN501, GLN498 and others residues preferably contemplate site also. In particular, the diversity of the virus in the world may be due to the genome structure of the virus and S gene changed over the time, across the world against to host of human genetic diversity, which may be more robust, and may be a new and unique feature. This is because it is characterized close to contact with distance divergence between wild type novel coronavirus which was risen from China against to the genomes from Lebanon, India, Italy, and USA and so on. Thus, the World Health Organization and its researchers should focus on immunologic research and effective drug and vaccine development that will help to address the epidemiology of the virus, which can provide a long-term solution.

Genetic Characteristics of <i>Citrus Tristeza Virus</i>Isolates from Cultivated Citrus in China Based on Coat Protein Gene

Citrus tristeza virus (CTV) is an important citrus pathogen causing considerable economic loss to citrus production. Knowledge on genetic evolutionary of the CTV population in China remains limited. In this study, 1439 samples were collected from nine citrus-producing areas of China. The coat protein (CP) genes of CTV were amplified by RT-PCR, and sequenced to analyze the genetic evolution. Analysis of the base composition showed an AU preference pattern, with the GC content was lower than AU content. Nine CTV populations were clustered into one clade in neighbor-joining (NJ) tree, indicative of a close phylogenetic relationship among the populations in China. Analysis of molecular variation (AMOVA) revealed that 77.72% genetic variations of CTV populations were observed among populations, with an FST value of 0.223. The values of dN/dS and neutrality test of CP gene were ranged from 0.016 to 0.082 and -1.377 to 1.456, respectively, the results suggesting that all of nine CTV populations were relatively constantly maintained under purifying selection. Our study demonstrated the genetic characteristics and molecular evolution relationship of CTV populations in China, and provided a theoretical basis for scientific control of CTV.

Genetic polymorphisms of MAFK, encoding a small Maf protein, are associated with susceptibility to ulcerative colitis in Japan

AIM To investigate whether single nucleotide polymorphisms in maf protein K(MAFK), which encodes the MAFK, lead to increased susceptibility to ulcerative colitis in the Japanese population. METHODS This case control study examined the associations between MAFK single nucleotide polymorphisms(rs4268033 G>A, rs3735656 T>C and rs10226620 C>T) and ulcerative colitis susceptibility in 174 patients with ulcerative colitis(UC) cases, and 748 subjects without no lower abdominal symptoms, diarrhea or hematochezia(controls). In addition, as the second controls, we set 360 subjects, who have an irregular bowel movement without abnormal lower endoscopic findings(IBM controls).RESULTS The genotype frequency of rs4268033 AA and allelic frequency of the rs4268033 A allele were significantly higher in the UC cases than in both controls(P = 0.0005 and < 0.0001, P = 0.015 and 0.0027 vs controls and IBM controls, respectively). Logistic regression analysis after adjustment for age and gender showed that the rs4268033 AA and rs3735656 CC genotypes were significantly associated with susceptibility to UC development(OR = 2.63, 95%CI: 1.61-4.30, P = 0.0001 and OR = 1.81; 95%CI: 1.12-2.94, P = 0.015, respectively). Similar findings were observed by the comparison with IBM controls. In addition, the rs4268033 AA genotype was significantly associated with all phenotypes of UC except early onset. There was no significant association between rs10226620 and ulcerative colitis. CONCLUSION Our results provide the first evidence that MAFK genetic polymorphisms are significantly associated with susceptibility to UC development. In particular, rs4268033 is closely associated with an increased risk for the development of UC.

Investigation of Aegilops umbellulata for stripe rust resistance,heading date,and the contents of iron,zinc,and gluten protein

Aegilops umbellulata(UU)is a wheat wild relative that has potential use in the genetic improvement of wheat.In this study,46 Ae.umbellulata accessions were investigated for stripe rust resistance,heading date(HD),and the contents of iron(Fe),zinc(Zn),and seed gluten proteins.Forty-two of the accessions were classified as resistant to stripe rust,while the other four accessions were classified as susceptible to stripe rust in four environments.The average HD of Ae.umbellulata was significantly longer than that of three common wheat cultivars(180.9 d vs.137.0 d),with the exception of PI226500(138.9 d).The Ae.umbellulata accessions also showed high variability in Fe(69.74-348.09 mg kg^(-1))and Zn(49.83-101.65 mg kg^(-1))contents.Three accessions(viz.,PI542362,PI542363,and PI554399)showed relatively higher Fe(230.96-348.09 mg kg^(-1))and Zn(92.46-101.65 mg kg^(-1))contents than the others.The Fe content of Ae.umbellulata was similar to those of Ae.comosa and Ae.markgrafii but higher than those of Ae.tauschii and common wheat.Aegilops umbellulata showed a higher Zn content than Ae.tauschii,Ae.comosa,and common wheat,but a lower content than Ae.markgrafii.Furthermore,Ae.umbellulata had the highest proportion of γ-gliadin among all the species investigated(Ae.umbellulata vs.other species=mean 72.11%vs.49.37%;range:55.33-86.99%vs.29.60-67.91%).These results demonstrated that Ae.umbellulata exhibits great diversity in the investigated traits,so it can provide a potential gene pool for the genetic improvement of these traits in wheat.

生物合成法生产麦角硫因的研究进展

麦角硫因(ergothioneine,ERG)作为一种稀有的天然含硫组氨酸衍生物,已被证明具有强大的抗氧化性和诸多生物学功能。因此,ERG受到越来越多研究人员和产品开发人员的关注。目前,ERG已被广泛应用于食品、化妆品和医疗等行业。研究表明只有少数细菌和真菌可体内合成ERG,植物、动物和人类自身均不能直接合成ERG,只能从其他来源获取。ERG可通过生物提取法、化学合成法以及生物合成法获得,但由于传统生产方式(生物提取法和化学合成法)存在产量低、生产效率差和生产成本较高等问题,限制了该产品的规模化生产和应用。因此,亟需开发一种高效、经济且安全、可靠的ERG生产方式以满足市场的需求。近年来合成生物学快速发展,利用基因工程、蛋白质工程和代谢工程等技术提高生物合成法生产ERG的能力已逐渐成为研究热点。本文将论述ERG的生物学活性和功能,介绍ERG生物合成途径和ERG在食品、化妆品和医疗等行业的应用前景,比较ERG主要的生产方式,总结并梳理近年来采取各种工程策略通过生物合成法生产ERG的研究进展;并就如何利用基因工程、蛋白质工程和代谢工程提高ERG产量提出几点工程策略,以期为生物合成法高产ERG提供理论参考和研究思路。

赤眼鳟LGP2序列结构、组织表达及与MDA5互作特征

为探究赤眼鳟遗传学和生理学实验室蛋白2(laboratory of genetics and physiology 2,LGP2)的功能特征及抗草鱼呼肠孤病毒(grass carp reovirus,GCRV)育种参考潜力,实验克隆获得了2940 bp的赤眼鳟lgp2(Sclgp2)全长cDNA和721 bp的5′端上游序列。Sclgp2 cDNA编码680个氨基酸,包含DEXDc(DExD/H-box helicase domain)、HELICc(helicase superfamily C-terminal domain)和CTD(C-terminal regulatory domain)结构域;其5′端上游序列含有MafB(muscle aponeurosis fibromatosis B)和IRF3(interferon regulatory factor 3)等转录因子结合位点。不同物种LGP2的功能结构域、磷酸化修饰位点数具有相似性,同时也存在结构域排布位置及序列的差异。赤眼鳟和草鱼lgp2 cDNA序列比较初步发现2个位于RNA结合功能区的GCRV抗性关联位点。系统进化分析显示,赤眼鳟LGP2先与草鱼、鲫和青鱼聚在一起,再与鲤科鱼类等聚为一大支。荧光定量表达分析显示,赤眼鳟脾脏中sclgp2表达水平显著高于其他组织,肌肉、心脏中表达量次之,而肠中表达量最低。GCRV感染后,肝脏中ifn1表达水平在24~72 h显著下降,其他组织sclgp2和ifn1表达水平未有显著变化。相关性分析结果显示,赤眼鳟肌肉sclgp2与ifn1表达水平呈极显著正相关(0.999)。酵母双杂交互作检测发现,赤眼鳟LGP2与MDA5存在弱相互作用,而其DEXDc(1~201 aa)、HELICc(390~476 aa)以及CTD(553~668 aa)结构域与MDA5无互作。该研究成功获得了sclgp2全长cDNA及5′端上游序列,明确了其序列结构、免疫表达及与MDA5的互作特征,为赤眼鳟LGP2免疫功能属性研究奠定了基础,并为草鱼抗GCRV育种提供了参考。

狂犬病病毒GX074株P-M^(S20F)突变体构建及其生长特性鉴定

【目的】明确狂犬病病毒(RABV)强毒株GX074的P蛋白和M蛋白第20位苯丙氨酸(Phe)替换至弱毒株r RC-HL的重组突变体在宿主细胞内的生长特性,为P蛋白和M蛋白转录复制机理研究提供理论依据。【方法】运用反向遗传技术以强毒株GX074的M蛋白第20位Phe替换弱毒株r RC-HL(GX074P)的M蛋白第20位丝氨酸(Ser),构建r RC-HL(GX074P-M^(S20F))突变体感染性c DNA克隆并拯救病毒。以重组突变体感染BSR/T7-9细胞,进行多步生长曲线测定,比较重组突变体与亲本毒株的生长能力,采用Western blotting检测N蛋白、P蛋白和M蛋白相对表达水平,利用实时荧光定量PCR检测N基因、P基因和M基因的相对表达量。【结果】经RT-PCR和测序鉴定,拯救的突变体r RC-HL(GX074P-M^(S20F))M蛋白第20位Ser成功替换为Phe。多步生长曲线测定结果显示,构建的重组突变体在感染24、48、72和96 h后,病毒滴度均高于亲本弱毒株r RC-HL和对照弱毒株r RC-HL(GX074PM1),平均约为亲本弱毒株r RC-HL的10倍。在蛋白表达水平上,r RC-HL(GX074P-M^(S20F))毒株在感染48 h后,N蛋白和M蛋白相对表达水平均极显著高于亲本弱毒株r RC-HL(P<0.01),说明强毒株GX074的P蛋白联合M蛋白第20位Phe替换至弱毒株r RC-HL后,增加了突变体N蛋白和M蛋白表达。感染24和48 h后,r RC-HL(GX074P-M^(S20F))毒株N基因、P基因和M基因的相对表达量均高于亲本弱毒株r RC-HL和对照弱毒株r RC-HL(GX074PM1)。【结论】强毒株GX074的M蛋白第20位Phe替换可提高病毒的增殖能力,同时增强病毒的复制与转录能力,M蛋白第20位Phe可能是影响病毒复制和转录的关键位点。

110份鹰嘴豆种质品质性状遗传多样性分析与综合评价

对国家中亚特色种质资源中期库(乌鲁木齐)保存的110份鹰嘴豆的7个品质性状进行遗传多样性分析与评价,筛选优异种质,为鹰嘴豆种质资源的挖掘利用和育种工作提供依据。结果表明,供试材料具有丰富的遗传多样性,7个品质性状变异系数变化范围为4.038%~23.950%,饱和脂肪酸含量变异最大,粗脂肪含量变异最小;Shannon-Weaver遗传多样性指数H′变化范围为1.205~2.040,淀粉含量的遗传多样性指数最高,饱和脂肪酸含量的遗传多样性指数最低。主成分分析将7个品质性状综合为4个主成分,累计贡献率为78.779%。聚类分析将110份鹰嘴豆种质资源划分为4个类群,其中第Ⅲ类群综合表现最好,该类群蛋白质、粗脂肪、淀粉、可溶性固形物和多不饱和脂肪酸含量均较高。综合评分F值范围为0.383~10.965,其中Desi类型和Kabuli类型的鹰嘴豆F值均值分别为7.396和7.175,根据F值筛选出5份综合表现好的品种,分别为‘品鹰1号’‘木鹰1号’‘2009-132’‘诺胡提’和‘引K158B’。

以痫性发作为表现成人起病的瓜氨酸血症Ⅱ型1例

对SLC25A13基因纯合变异引起以痫性发作为表现的成人起病的瓜氨酸血症Ⅱ型(adult-onset typeⅡcitrullinemia,CTLN2)1例进行回顾性分析。患者,男,28岁,反复四肢抽搐4年余,再发加重2个月,平素喜食花生及肉类。头颅MRI检查未见异常,予抗癫痫治疗效果不佳,进一步查血转氨酶、血氨和瓜氨酸升高,基因检测显示SLC25A13基因c.851_854del纯合致病突变,诊断为CTLN2,予高蛋白、高脂肪、低糖饮食和精氨酸治疗,随访半年无痫性发作。对反复痫性发作伴有特殊饮食嗜好者应注意CTLN2可能,基因检测对CTLN2的诊断具有重要作用,可为临床诊治提供依据。

体外膜氧合联合介入取栓救治蛋白C基因突变所致高危肺栓塞一例

遗传性蛋白C缺陷症是由蛋白C基因突变引起的一种染色体遗传病,可导致静脉血栓形成,多与外显子4~9和内含子8的突变有关。蛋白C基因突变引起的致死性肺栓塞罕见,治疗面临巨大挑战。本文报道1例由蛋白C基因8号外显子移码突变引起的致死性肺栓塞,采用体外膜氧合进行呼吸、循环支持,并成功实行介入取栓的救治经验,为该疾病的诊断及救治提供参考。