小麦RIL群体中籽粒蛋白质组分的分离与主要品质性状的相关分析

利用普通小麦重组自交系群体为材料,研究了小麦籽粒蛋白质组分在后代群体中的分离和分布,同时对籽粒蛋白质组分与其它主要品质指标的关系进行了研究.结果发现籽粒蛋白质组分在群体后代中表现为微效多基因控制的数量性状,相对亲本的性状变异较大.醇溶蛋白和谷蛋白相对含量与湿面筋含量、SDS沉降值之间呈显著正相关,与硬度呈正相关.清蛋白、球蛋白相对含量与湿面筋含量、SDS沉降值和硬度呈负相关.     

一个在梅山猪×大白猪杂交组合的背最长肌中差异表达的新基因的分离、cDNA 序列分析及组织表达谱

为了揭示猪杂种优势的分子机理,用mRNA差异显示技术从梅山猪×大白猪杂交组合的背最长肌中分离到一个差异表达的基因,并用半定量RT-PCR进行鉴定,随后通过cDNA末端快速扩增法(RACE)得到该基因的cDNA全长.经与GenBank进行Blast比较,这个基因与猪的已知的基因没有明显的同源性.基因预测显示这个基因编码一个由188个氨基酸组成的蛋白质,且该蛋白质具有保守的PRA1 family protein的结构域,同时该蛋白质和人、大鼠、小鼠的PRA1 family protein 3分别具有88%,88%,87%的同源性,因此将这个基因命名为猪的PRA1 family protein 3基因.进化树分析表明猪的PRA1family protein 3和人的PRA1family protein 3具有比大鼠、小鼠的PRA1family protein 3更近的亲缘关系.组织表达谱分析显示这个基因在肌肉与脂肪组织中表达丰富,在脾中中等表达,在心、肾、卵巢、肺中少量表达,在肝与小肠中几乎不表达.同时对这个基因的功能及与猪的杂种优势的关系进行了讨论.     

蛋白质二级结构中同义密码子与氨基酸上下文关联的偏好型

通过对人(Homo sapiens)的117个蛋白质和大肠杆菌(E．coli)的87个蛋白质的统计分析，发现mRNA序列中的同义密码子与氨基酸的上下文关联是和蛋白质二级结构有关的，并给出了各二级结构中有意义的上下文关联型．讨论了该结果对蛋白质结构预测的改进意义及其在基因工程领域可能的应用。     

Inhibitory effect of the adenovirus type 5 E1A protein expressed in the yeast system

The human adenovirus type 5 E1A, a tumor- suppressor gene[1], codes for two major related proteins of 243 amino acids (12S) and 289 amino acids (13S) by al-ternative splicing in two exons[2]. Studies have been shown that E1A can regulate expression of many genes and cell cycle[3]. Both in vitro and in vivo experiments indicated that E1A could induce tumor cells differentia-tion, convert tumor cells into an epithelial phenotype, in-hibit tumor cell growth and metastasis and strongly en-ha…     

Roles of protein kinase C in oocyte meiotic maturation and fertilization

Protein kinase C (PKC) is a superfamily of Ser/Thr protein kinases that is distributed widely in eukaryotes. It plays key regulatory roles at multiple steps of oocyte meiotic maturation and fertilization. During the process of meiotic maturation, the activation of PKC in cumulus cells stimulates meiotic maturation, whereas the activation of PKC in oocytes results in the inhibition of germinal vesicle breakdown. PKC activity increases following the meiotic maturation, and decreases at the transition of metaphase/anaphase in meiosis I, so as to facilitate the release of the first polar body and the entry of meiosis II. In fertilization of mammalian oocytes, PKC may act as one of the downstream targets of Ca2+ to stimulate the cortical granule exocytosis, release the oocytes from MII arrest and to induce pronucleus formation. PKC is also involved in the regulation of maturation promoting factor (MPF) and mitogen-activated protein kinase (MAPK). Several PKC isoforms have been identified in mammalian oocytes, and there is evidence showing that classical PKCs may be the principal mediator of oocyte cortical reaction.     

N-terminal of L protein of vesicular stomatitis virus contains a new signal sequence

The L protein (241kD) of vesicular stomatitis virus (VSV) is the most important snbnnit of the replication complex. The existence of specific localization signal in the L protein was investigated by making recombinant constructs expressing truncated mutants of the L protein fused to green fluorescent protein (GFP) in transient transfection assays. The chimeric genes encoding varied N-terminal of L and GFP gene were put under the control of T7 promoter or CMV promoter. The fusion proteins were transiently expressed in BHK-21, COS-7, CHO or Hep G2 cells. When more than 120 residues were deleted or only 96 residues were kept on the N-terminal, the fusion proteins were shown to be distributed throughout the cells, cytoplasm and nucleus under the confocal microscope. However, other chimeric proteins with 120 or more amino acids were dotted and distributed in the perinuclear regions. And the fusion protein with 96—120 aa has the similar distribution. A thirteen-residue peptide QGYSFLHEVDKEA (108—120) was identified as localization signal, whose function would be absolutely distributed with the deficiency of D or V. Our results show that there is an independent localizing signal in N-terminal domain of L protein of VSV and this functional signal is conserved in different cell lines.     

Analysis of cDNA sequence, protein structure and expression of parotid secretory protein in pig

Parotid secretory protein (PSP) secreted abundantly in saliva, whose function is related with the anti-bacterial effect. The PSP cDNA has been isolated from pig parotid glands by 3′ and 5′ rapid amplification of cDNA end (RACE), based on the conserved signal peptide region among the known mammalian PSP. The result of homologous comparison shows that pig PSP and human PSP shares the high identity at the level of the primary, secondary and tertiary protein structure. A search for functionally significant protein motifs revealed a unique amino acid sequence pattern consisting of the residues Leu-X(6)-Leu-X(6)-Leu-X(7)-Leu-X(6)-Leu-X(6)-Leu near the amino-terminal portion of the protein, which is important to its function. RT-PCR, Dot blot and Northern blot analysis demonstrated that PSP was strongly expressed in parotid glands, but not in other tissues.     

Inhibitory effect of the adenovirus type 5 E1A protein expressed in the yeast system of the human tumor cell growth

Adenovirus 5 type E1A as a tumor suppressor gene can inhibit tumor growth and enhance the censitivity of chemotherapy and radiotherapy.E1A have the ability to integrate into the host genome,resulting in long-time expres-sion that induces Rb gene inactivation and animal cells im-mortalization.This prompted us to select the E1A protein for treatment of cancer in order to overcome the limitations of E1A gene therapy.Thus,we firstly comstructed E1A eu-caryotic expression vector (pPIC9/E1A),transformated the pichia pastoris yeast cells(GS115) and screened the high-expressing recombinant strains.The positive yeast strains were cultured in the shake flask,and induced for 3d.The crude E1A protein was purified using two steps of col-umu chromatography on HiTrap Q and HiTrap SP.The pu-rified E1A protein was identified by SDS-PAGE and Western blot.E1A protein was mostly located at cellular unclear when Cheriot delivered E1A protein into cells.The analysis in vitro indicated that the E1A protein arrested LN686 cell cycle at G2/M phase,and significantly inhibited the growth of LN686 tumor cells.The current studies firstly provided an experimental basis to further develop E1A protein for tumor treatment.