青山羊小肠凝集素的分离,纯化及性质研究

报道了青山羊小肠凝集素的分离,纯化及性质研究。青山羊小肠先经过含有疏基乙醇的磷酸缓冲液抽提,然后上Ｓｅｐｈａｒｏｓｅ ６Ｂ柱及ＤＥＡＥ－ＣＥＬＬＵＬＯＳＥ－２３柱,得到纯化的青山羊小肠凝集素,采用ＳＤＳ电泳法测得其分子量在６６ １００左右,而且该凝集素不含糖,对人Ｂ型血球有专一性凝集作用。半抗原抑制实验表明它对半乳糖（乳糖）有亲和性,其中酸性氨基酸含量较高,组氨酸,蛋氨酸含量较低,该凝集素在胚胎期     

苦参凝集素的分离纯化及部分性质研究

从苦参 (SophoraflavescensAit.)根浸出液经硫酸铵分级 ,得苦参凝集素 (SFL)粗品 ,再经DEAE_Sepharose、SephadexG_1 5 0和HPLC层析 ,获得具有强凝集活性的SFL样品 ,用PAGE和SDS_PAGE检测均为单一蛋白染色带。SDS_PAGE显示SFL分子仅有一条肽链 ,SephadexG_1 0 0和SDS_PAGE测得其分子量均为 32kD。当SFL浓度为 0 .97μg/mL时能凝集兔红细胞 ,无血型专一性 ,其凝血活性可被甘露糖和果糖抑制 ,麦芽糖和葡萄糖有弱的抑制作用 ,凝集素分子含有 2 .89%的中性糖 ;当SFL量为 6 2 μg时 ,对棉花枯萎病菌 (FusariumvasinfectumAtk .)、小麦赤霉病菌(Gibberllasaubinetii (Mont.)Sacc .)和水稻稻瘟病菌 (PiriculariaoryzaeCav)菌丝体的生长发育有明显地抑制作用。用Edman法在蛋白测序仪上测出SFL的N端肽链 30个氨基酸的排列顺序为 :T/A/VDXLXFTFSDFDP NGEDLLFQGDAHVTSNN。     

江苏菜豆同工凝集素的分离纯化及性质研究

江苏菜豆经酸水(PH2.0)抽提,硫酸铵分级沉淀,分离植物血球凝集素(PHA-P),分子量为128000的糖蛋白,活性回收率在80％以上,PHA-P经SP-sephadexc-50离子交换层析,分成L_4,L_3E_1,L_2E_2,L_1E_3,和E_4同工凝集素。 L_4和E_4等电点为5.4和6.5。亚基分子量分别是31000和33000,并有类似的氨基酸组成。PAGE分析为单一蛋白带。红细胞凝集活性随电泳迁移速度的加快而增强,促淋性细胞分裂活性则减弱。E_4血凝活性受CalNAc,EDTA抑制和Zn~(++)的促进。     

中国鲎凝集素的分离纯化及性质研究

 经N-乙酰氨基葡萄糖交联琼脂糖亲和层析及以交联琼脂糖介质的高效液相分子筛层析,从中国鲎细胞溶解物中分离纯化了一种凝集素,其活性比原料鲎试剂提高128倍。鲎凝集素SDS电泳时表现出分子量为69000,和72000的二个亚基。N-乙酰氨基葡萄糖、D-半乳糖,D-甘露糖及岩藻糖等对鲎凝集素凝集鸡红细胞的活性有显著抑制作用,加热60℃,10分钟可使凝集素活性基本丧失。CaCl_2为凝集素活性所必需。鲎凝集素与肺炎球菌C多糖有沉淀反应。     

桑寄生凝集素的纯化及部分性质研究

利用DEAE一纤维素柱盐离子浓度梯度洗脱,再经猪胃粘蛋白-Sepharose 4 B亲和柱可以从中药桑寄生中分离纯化出桑寄生凝集素。经pH8.9,Tris-EDTAN_2a-borate的PACE和SDS-PAGE测定均呈现单一蛋白带,测得其分子量为67 500,中性糖含量为14.6％,DNS法测得N-末端氨基酸为缬氨酸。氨基酸组成分析表明,该凝集素富含酸性氨基酸,而碱性氨基酸含量较少,不含精氨酸。当凝集素浓度为15.6μg/mL时,即可凝集兔红细胞,而对人的A、B、O型血细胞,凝集素浓度高达1000μg/mL,也不发生凝集反应。Gal、GalNAc、山梨糖、岩藻糖和松三糖对凝集兔红细胞的能力有抑制作用。桑寄生凝集素是一种促有丝分裂原,对猪血淋巴细胞的转化率达78％,细胞分裂比率为11.2％。     

紫藤凝集素的分离纯化及理化性质研究

用常规方法处理的ＤＥＡＥ离子交换纤维素柱,通过线性离子强度梯度洗脱,从紫藤种子的蛋白粗提液中得到一定纯度的紫藤凝集素。纯化的凝集素凝集兔红血球的比活提高４０倍,总活力回收率为１９．２％。紫藤凝集素的分子量经ＰＡＧＥ鉴定为２０５ｋｄ,是由两种亚基构成的四聚体,这两种亚基各有２个,分子量ＳＤＳ－ＰＡＧＥ鉴定分别为７７６００ｄ和２５１００ｄ。紫藤凝集素是一种糖蛋白,等电点约为４．６０。它可凝集人的各种血     

Eimeria Caprovina Sp. N. From the Domestic Goat, Capra Hircus, From the Usa

Oocysts of Eimeria caprovina sp. n. from the domestic goat, Capra hircus , are ellipsoidal, subspherical or slightly ovoid, usually flattened at the micropylar end. They measure 29.7 (26-36) × 23.7 (23-28) μ. the sporocysts are elongate ovoids, measuring 14.3 (13-17) × 8.3 (8-9) μ. with Stieda bodies at the narrow ends. the oocyst wait is 1.6 μ thick, smooth, dark-brown to brownish-yellow, and 2-layered. A micropyle. 6.2 (4-10) μ in diameter, polar granule, and sporocyst residuum are present: micropylar cap and oocyst residuum are absent.     

Biochemical and genetic analysis of variant C of caprine αs2-casein (Capra hircus)

Two alleles, A and B, were previously described at the goat α_s2-casein locus. Isoelectric focusing allowed us to subdivide the former one in two new alleles, called A and C. Although α_s2-casein C cannot actually be distinguished from its A counterpart by starch or polyacrylamide gel electrophoresis, it differs from the previous allele by a single substitution Lys (A)/Ile (C) at position 167, which was confirmed at the nucleotide level. The frequencies of the three α_s2-casein alleles A, B and C were estimated to be 0.85, 0.04 and 0.11 in the French dairy breeds ‘Alpine’ and ‘Saanen’.     

An allele associated with a non-detectable amount of αs2 casein in goat milk

The goat CSN1S2 locus is characterized by the presence of three alleles, A, B and C, all associated with about 2.5 g/l of protein per allele. The SDS-PAGE analysis of 441 individual milk samples obtained from goats belonging to a population reared in Southern Italy showed that the milk produced by three goats did not apparently contain alpha s2-casein, whereas milk produced by 37 goats showed a less intense electrophoretic band of this casein fraction (about 50%). These results can be explained by hypothesizing the presence of another allele at this locus, CSN1S2o, associated with a 'null' content of alpha s2-casein. Southern blot, PCR and PCR-RFLP analyses of the DNA region containing the CSN1S2 gene of individuals producing milk with and without alpha s2-casein did not show differences between the two groups. As a consequence, goats producing milk without alpha s2-casein carry an apparently intact gene. The first results obtained by sequencing part of the CSN1S2o allele revealed a G-->A transition at nucleotide 80 of the 11th exon which creates a stop codon and could be responsible for the absence of the alpha s2-casein in goat milk. This mutation eliminates a NcoI restriction site. A test based on this polymorphism has been established in order to identify carriers of the CSN1S2o allele.     

Daily rhythm of body and auricle temperature in goats kept at two different ambient temperatures

In fluctuating environmental temperature, homeotherms are able to maintain stable their body temperature, which however reveals a rhythmic daily pattern as described in literature. Because of the importance of body temperature rhythmicity in the knowledge of thermal homeostasis and as a means to facilitate the study of biological rhythmicity in general, the aim of our study was to assess the influence of two different ambient temperature on the daily rhythmicity of body and auricle temperature in goat (Capra hircus). For our study 6 female adult Maltese goats (18 month old), not pregnant and clinically healthy were used. On each subject body and auricle temperature was recorded during two different periods, of 7 days each, every 3 hours for 24 hours from 07.00 to 04.00, at different environmental temperature (1^st period 18.25 ± 1.48°C, 2^nd period 28.25 ± 2.05°C). The analysis of the obtained result under two ambient temperature indicates the existence of a daily periodicity, with small differences in amplitude and acrophase between the two studied period, both in body and auricle temperature in Maltese breed goat so we can assume that environmental temperature can influence in a significant way the daily pattern of body and auricle temperature.     

Effects of Nu-Serum® on in vitro development of goat (Capra hircus) embryos

To evaluate the effects of a commercially produced serum substitute on the in vitro development of caprine embryos, registered Nubian doelings were synchronized with norgestomet-impregnated implants (Synchromate-B®: CEVA) and superovulated with descending doses of FSH-p® (Schering). A total of 246 embryos was collected and placed in Tissue Culture Medium 199 (TCM 199, Gibco Laboratories) containing Nu-Serum® (NuS) at concentrations of 2.5%, 5.0%, 10%, or 20%. Control treatments consisted of TCM 199 alone or TCM 199 plus 10% heat-inactivated fetal bovine serum (FBS). Embryos developed in all concentrations of NuS to the morula, blastocyst, and expanded blastocyst stages. The TCM 199 plus 10% NuS had significantly higher percentages of embryos developing to the expanded blastocyst stage than TCM 199 plus 10% FBS. Time to expanded blastocyst development in NuS was shorter than in the TCM 199 plus FBS. No stage-specific block to development was observed with embryos collected and cultured in vitro for any of the treatments. These results demonstrate that NuS, when compared to FBS, allowed a higher percentage (P < 0.05) of caprine embryos to develop to the expanded blastocyst stage, thus providing a valuable substitute for FBS.     

Characterization of two new alleles at the goat CSN1S2 locus

Two novel alleles at the goat CSN1S2 locus have been identified: CSN1S2(F) and CSN1S2(D). Sequence analyses revealed that the CSN1S2(F) allele is characterized by a G --> A transition at the 13th nucleotide in exon 3 changing the seventh amino acid of the mature protein from Val to Ile. The CSN1S2(D) allele, apparently associated with a decreased synthesis of alpha s2-casein, is characterized by a 106-bp deletion, involving the last 11 bp of the exon 11 and the first 95 bp of the following intron. Methods (PCR-RFLP and PCR) for identification of carriers of these alleles have been developed.     

Geographical partitioning of goat diversity in Europe and the Middle East

Thirty microsatellite markers were analysed in 1426 goats from 45 traditional or rare breeds in 15 European and Middle Eastern countries. In all populations inbreeding was indicated by heterozygosity deficiency (mean FIS = 0.10). Genetic differentiation between breeds was moderate with a mean FST value of 0.07, but for most (c. 71%) northern and central European breeds, individuals could be assigned to their breeds with a success rate of more than 80%. Bayesian-based clustering analysis of allele frequencies and multivariate analysis revealed at least four discrete clusters: eastern Mediterranean (Middle East), central Mediterranean, western Mediterranean and central/northern Europe. About 41% of the genetic variability among the breeds could be explained by their geographical origin. A decrease in genetic diversity from the south-east to the north-west was accompanied by an increase in the level of differentiation at the breed level. These observations support the hypothesis that domestic livestock migrated from the Middle East towards western and northern Europe and indicate that breed formation was more systematic in north-central Europe than in the Middle East. We propose that breed differentiation and molecular diversity are independent criteria for conservation.