乳腺特异表达载体缺失片段的鉴定与修复

通过对pBC1中的乳腺特异表达启动子成分进行分析,鉴定了位于β酪蛋白启动子上游两个EcoRⅠ位点(-89~-94和-120~-125)之间的缺失片段,31bp的缺失使得该载体中的一个乳控盒元件(-112~-141)和一个乳腺因子识别序列(-92~-102)遭到破坏,并使得位于TATA框上游的所有启动子成分发生了位置改变。通过与山羊β酪蛋白启动子序列进行比较,设计并合成了缺失片段,经一系列的酶切和连接处理,使得该缺失得到了修复,修复后的载体具有所有乳蛋白表达需要的启动子调控原件。本试验结果为利用修复后的乳腺特异高效表达载体进行乳腺生物反应器的研究奠定了分子生物学基础。     

军牧1号白猪、杜洛克猪和藏猪的氟烷基因和酸肉基因多态性分布

为了研究军牧1号白猪、杜洛克猪和藏猪氟烷基因和酸肉基因的多态性分布情况,试验采用PCR-RFLP方法对61头军牧1号白猪、51头杜洛克猪和51头藏猪的氟烷基因和酸肉基因多态性进行了检测。结果表明:军牧1号白猪的氟烷基因表现为单一的NN型;杜洛克猪的氟烷基因表现为多态性,等位基因N的频率为0.196 1,基因型分布符合哈代-温伯格平衡(χ2=3.033 9,P=0.081 5);藏猪的氟烷基因表现为单一的nn型。3个猪种的酸肉基因均表现为单一的rn/rn型。     

Q型烟粉虱芳香基硫酸酯酶B基因的克隆及表达分析

为明确烟粉虱Bemisia tabaci取食感染番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)的番茄植株后,其体内的芳香基硫酸酯酶B基因(arylsulfatase B,ARSB)是否能够做出应答反应,基于Q型烟粉虱基因组数据克隆得到ARSB基因cDNA全长,采用生物信息学方法分析其序列特征,并通过实时荧光定量PCR技术测定ARSB基因在Q型烟粉虱不同发育阶段、不同组织及携带TYLCV前后的表达量变化情况。结果显示:Q型烟粉虱ARSB基因的cDNA全长为1 731 bp,编码576个氨基酸,分子量为64.89 kD,具有ARSB的保守结构域。ARSB基因在Q型烟粉虱不同发育阶段均有表达,在卵期表达量最高,成虫期表达量最低;该基因在Q型烟粉虱头胸部的表达量显著高于腹部;Q型烟粉虱获取TYLCV 72 h后其体内ARSB基因表达量显著提高。表明ARSB基因在Q型烟粉虱不同龄期、不同组织内存在差异表达,并可能参与Q型烟粉虱对TYLCV的响应和传毒过程。     

Cloning of Proteinase Inhibitor Gene StPI in Diploid Potato and Its Expression Analysis

A full-length cDNA of proteinase inhibitor gene with completed open reading frame of 116 amino acids was cloned from Ralstonia solanacearum (Rs) resistant potato leaves using the rapid amplification of cDNA ends (RACE) method and designated as StPI. BLAST search against NCBI showed that the StPI gene shared 89% identity with potato proteinase inhibitor Ⅰ precursor in nucleotide and 74% in amino acid. Analysis of semi-quantitative RT-PCR indicated that this gene was induced by Rs as well as up-regulated by jasmonic acid (JA). The StPI gene expression reached the highest level during 6-12 h post Rs-inoculation or JA-treatment, and then leveled off. Moreover, this gene was strongly induced by JA and its mRNA accumulation increased more quickly than that of Rs-inoculation. The StPI gene may play a role in potato resistance against Rs. The induction of StPI by Rs invasion may have a similar signal transduction pathway with JA treatment.     

不同物种CLPG基因部分序列生物信息分析及山羊染色体电子定位

作者应用比较基因组学和生物信息学方法比较了绵羊、山羊、牛、狗、马、兔子、猪、挪威鼠的CLPG基因部分序列，并对该基因的遗传多样性和遗传分化进行了分析。结果表明，在被比对的12个基因序列中检测到176个多态位点，物种间及物种内的CLPG基因存在较丰富的遗传多样性。通过物种间生物信息分析将CLPG基因定位于山羊21号染色体。     

Characterization of MC1R and KIT Genotypes in Luchuan Pig Population

In order to assess the purity of Luchuan pig populations, four South Chinese local pig breeds including Putian Black pig, Yuedong Black pig, Dahuabai pig, Bama miniature pig and three foreign pig breeds including Duroc pig, Yorkshire pig and Landrace pig were studied as controls by sequencing of PCR products, MC1R and KIT genotypes in 56 Luchuan pigs were analyzed in this study. Sequencing results indicated that a splicing mutation (G>A) was presented in the first base in intron 17 of KIT gene in both Yorkshire pig and Landrace pig, in contrast, the wildtype GG of KIT gene was presented in Luchuan pig, four south Chinese local pig breeds and Duroc pig.Compared with Hainan wild boar, South Chinese local pig breeds had two missense mutations 95Val > Met and 102Leu > Pro in the coding region of MC1R gene;Compared with Yorkshire pig, Landrace pig and Duroc pig, South Chinese local pig breeds had 5 to 6 SNPs in MC1R gene 5'UTR, and in addtion, an A base deletion in MC1R gene 3'UTR. Furthermore, we found one litter of Luchuan pig with abnormal coat color.The results showed that the presentation of two distinct MC1R genotypes E^D1 and E^p in both litters and the sow,but only E^D1 in the boar. Considering E^p was derived from Pietrain pig, we preliminarily considered that the genome of the sow might be infiltrated with foreign pig breeds. In summary, we detected the genotypes of the coat color genes KIT and MC1R in eight pig breeds, confirmed the molecular differences of coat color between Chinese local pig breeds and foreign pig breeds, which could be useful for the further investigation of the molecular mechanism of pig coat color.     

辣椒GRAS家族全基因组鉴定与表达分析

基于辣椒全基因组数据信息,利用生物信息学方法对CaGRAS基因家族进行了系统鉴定和进化分析,并通过Real-time PCR方法检测了CaGRAS家族组织表达模式和对PEG6000、盐胁迫的应答。结果表明:在辣椒中存在54个CaGRAS基因,除11号染色体外其余染色体均有分布,外显子数1~3,等电点4.97~9.13;系统进化分析显示CaGRAS基因可分为8个进化群;CaGRAS基因具有不同的表达模式,在根、茎、叶片和茎尖中优势表达的基因分别有34、8、3和8个;多数CaGRAS基因能响应PEG6000和盐胁迫,其中CaGRAS11、CaGRAS30、CaGRAS40、CaGRAS44和CaGRAS50受到PEG6000和盐胁迫的强烈诱导。本研究为深入解析CaGRAS家族基因的功能奠定了一定基础。     

Differential Expression Analysis of MUC4 and MUC13 Genes Between Resistant and Sensitive Weaned Meishan Piglets to ETEC F18

MUC4 and MUC13 genes as important candidate genes for enterotoxigenic Escherichia coil (ETEC) F4 resistance,may play an important role in the process of against ETEC F18 infection in weaned piglets. In this study,ETEC F18-resistant and -sensitive weaned Meishan piglets were used,and the expression levels of MUC4 and MUC13 genes in 11 tissues (heart,liver,spleen,lung,kidney,stomach,muscle,thymus,lymph nodes,duodenum and jejunum) were determined by quantitative Real-time PCR. The results showed that MUC4 and MUC13 genes were broadly expressed with different expression levels in all the 11 tissues. In the thymus and lymph tissues,the expression of MUC4 gene in resistant piglets was significantly higher than that in sensitive piglets (P<0.05);In the lung tissue,theMUC13 gene expression level in resistant individuals was significantly higher than that in sensitive individuals (P<0.05),and in the intestinal tissues of duodenum and jejunum, the expression level of MUC13 gene was relatively higher in resistant individuals. Thus we speculated that the high expression of MUC4 gene in immune tissues and MUC13 gene in intestinal tissues might improve the immune ability of piglets,protect and lubricate the intestinal tract, and resist ETEC F18 infection.