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1.
猪生长激素脂质体对育肥猪缓释作用效果研究   总被引：1，自引：0，他引：1  
为了解猪生长激素(pGH)脂质体对育肥猪缓释作用效果,选用体重约50 kg健康去势的长白×大约克公猪24头,随机分成4组,即对照组和3个试验组。对照组每天注射生理盐水,试验一组每天按4 mg的剂量肌肉注射pGH,试验二组每3 d注射12 mg pGH脂质体,试验三组每7 d注射28 mg pGH脂质体,试验期为21 d。结果显示:注射pGH及其脂质体能够提高猪的日增重(P<0.05)和末重,显著降低料重比(P<0.05),试验组血清中胰岛素样生长因子1(IGF-1)水平显著高于对照组(P<0.01);注射生长激素的猪血清中生长激素水平均有不同程度的升高。脂质体的缓释时间可以达到7 d,表明猪生长激素脂质体在促进猪生长发育方面的效果明显。  相似文献   

2.
猪生长激素基因在昆虫细胞中的分泌表达   总被引：9，自引：0，他引：9  
将PCR将增得到pGH基因插入到带有polh启动子和gp67强信号肽序列的杆状病毒转移载体pAcGP67-A中，构建重组质粒pGP67pGH，并与线性化致死缺失型苜蓿丫纹夜蛾核多角体病毒（AcMNPV-OCC)基因组DNA共转染Sf9细胞，构建出重组病毒AcMNPV-pGP67-pGH-OC^-。感染重组病毒的Hi5细胞的表达产物的SDS-PAGE和Western blot结果表明，细胞可溶蛋白和培养液上清中均有一条分子量约为22kDa的猪生长激素特异性反应带，且培养液上清中的目的蛋白分子量比天然pGH略大一些，薄层扫描仪扫描估测可知，重组pGH分别占细胞可溶蛋白的8．98％和培养液上清总蛋白的3.61%。将表达96h的培养液上清浓缩液进行N－糖基化分析，结果显示重组pGH无N－糖基化加工修饰。  相似文献   

3.
含第一内含子猪生长激素cDNA基因在COS7细胞中的表达   总被引：4，自引：1，他引：3  
以CMV真核生物启动子构建了含第一内含子猪生长激素（pGH）cDNA基因（pGHcDNA-in）的表达载体，经DEAE-葡聚糖介导在COS7细胞进行了瞬时表达，通过PP95生物学测定法测定，证明含第一内含子pGHcDNA基因能够在真核细胞中进行分泌性表达，表达出的pGH具有生物学活性。这说明通过该基因转录出的前体mRNA能够在COS7细胞中进行正确剪接，从而翻译出具有生物活性的pGH。加入第一内含子在一定程度上可提高pGHcDNA基因的表达效率。  相似文献   

4.
猪生长激素基因的克隆及在哺乳动物细胞中的表达   总被引：4，自引：0，他引：4  
应用RT—PCR技术克隆了猪生长激素(pGH)cDNA，该基因编码蛋白的信号肽序列与已有报道的pGH基因存在2个氨基酸残基的差异，而成熟肽却无差异。将pGHcDNA定向插入真核表达载体VRl020，构建了重组真核表达质粒VpGH；利用脂质体法转染哺乳动物细胞COS7，对转染后的COS7细胞进行RT—PCR、ELlSA和免疫荧光分析，分另1在转录和翻译水平证实了目的基因在COS7细胞中得到正确转染表达。  相似文献   

5.
猪生长激素基因的原核表达及其抗体制备   总被引：6，自引：0，他引：6  
通过构建pGH基因的原核表达质粒，转化大肠埃希氏菌TOP10，经IPTG诱导，成功表达了重组猪生长激素的融合蛋白。经SDSPAGE分析，在分子质量50．4ku处有1条新的特异蛋白质条带。对以包涵体形式表达的融合蛋白进行SDSPAGE分离，并经透析得到了重组融合蛋白，以此融合蛋白免疫家兔，制备并纯化了抗pGH多克隆抗体，经酶联免疫吸附测定、蛋白质印迹分析和免疫组织化学方法检测，此抗体具有较高效价和特异性。成功地表达了pGH基因并获得了多克隆抗体。  相似文献   

6.
为构建猪附红细胞体ENO基因重组腺病毒穿梭质粒,试验根据GenBank中登录的猪附红细胞体ENO基因序列（登录号：CP002525.1）设计特异性引物,对ENO基因进行PCR扩增,并将纯化后的PCR产物克隆到pMD19-T载体中。用Kpn Ⅰ和Xho Ⅰ对pMD19T-ENO进行双酶切后,将其亚克隆至腺病毒穿梭载体PCR259中,构建PCR259-ENO重组质粒,提取重组质粒进行鉴定。应用脂质体介导转染法将鉴定正确的PCR259-ENO重组穿梭质粒转染293细胞,应用间接免疫荧光法（IFTA）检测ENO基因在293细胞中的表达。结果显示,试验克隆的ENO基因长为1 182 bp,编码393个氨基酸,与GenBank中ENO基因序列（登录号：CP002525.1）同源性为99%。构建的重组腺病毒穿梭质粒PCR259-ENO经PCR和酶切鉴定正确,并且能在293细胞中表达,表明ENO基因成功插入腺病毒穿梭质粒PCR259中,重组腺病毒穿梭质粒PCR259-ENO构建成功。  相似文献   

7.
试验旨在构建表达猪附红细胞体ENO基因的重组腺病毒并分析评价其免疫效果。将重组克隆质粒pMD-19T-ENO与腺病毒穿梭载体AdV4-GFP分别进行双酶切,构建重组腺病毒穿梭质粒AdV4-M/ENO;将经PacⅠ酶线性化后的重组腺病毒穿梭质粒AdV4-M/ENO转染293细胞,获得重组腺病毒Ad4-M/ENO,采用PCR和间接免疫荧光试验（IFTA）鉴定猪附红细胞体ENO基因在293细胞中的表达,再对293细胞进行培养,测定重组腺病毒的滴度;将30只BALB/c小鼠分为3组：重组腺病毒Ad4-M/ENO组、AdV4-GFP空载体对照组和PBS对照组,分别进行免疫接种,采用ELISA方法检测血清中猪附红细胞体IgG、IgG1、IgG2a抗体水平和IFN-γ、IL-4细胞因子水平,在三免2周后检测小鼠脾脏中CD4+和CD8+含量。结果显示,构建的重组腺病毒穿梭质粒AdV4-M/ENO目的基因片段大小为1 182 bp;重组腺病毒Ad4-M/ENO包装成功,能在293细胞中表达,滴度为1×109 PFU/mL。经重组腺病毒Ad4-M/ENO免疫后的BALB/c小鼠血清中IgG、IgG1、IgG2a抗体水平,IFN-γ、IL-4细胞因子水平及淋巴细胞亚群CD4+、CD8+含量均显著或极显著高于AdV4-GFP空载体对照组和PBS对照组（P<0.05;P<0.01）。结果表明,本试验成功构建了表达猪附红细胞体ENO基因的重组腺病毒,且该重组腺病毒能诱导小鼠产生特异性的体液免疫和细胞免疫应答反应。  相似文献   

8.
猪生长激素的研究进展   总被引：1，自引：0，他引：1  
猪生长激素(porcinegrowthhormone,pGH)是一种由猪脑垂体前叶嗜酸性细胞分泌的单一肽链蛋白质类激素。1920年,Evans和Simpson给大鼠注射牛垂体提取物,首次证实GH具有促生长作用,因此称之为生长激素。在生长轴中,GH是调控动物生长发育的核心,是一种具有广泛生理功能的生长调  相似文献   

9.
应用RT-PCR方法扩增出猪日本乙型脑炎病毒（JEV）的NS1基因,通过Sal Ⅰ＋EcoR Ⅰ双酶切后把该基因插入到经过同样双酶切的穿梭质粒pDC315中.重组穿梭载体经过双酶切和PCR鉴定后进行测序,序列测定正确.将获得的重组穿梭质粒与腺病毒骨架质粒共转染293细胞后获得重组腺病毒.PCR鉴定及间接ELISA检测的结果证明所构建的重组腺病毒成功地表达了JEV的NS1.  相似文献   

10.
生长速度和胴体品质是养猪生产过程中2个重要的经济性状。猪生长激素(pGH)是一种具有广泛生理功能的生长调节素,其主要作用是通过改善营养物质在猪体内的配比,促进蛋白质合成及抑制脂肪生成,进而提高动物体增质量和瘦肉率。  相似文献   

11.
Twenty-eight barrows were used to investigate the effects of exogenous porcine growth hormone (pGH) administration (0 and 100 micrograms.kg-1.d-1) between 30 and 60 kg on the subsequent and overall performance and carcass composition of pigs grown to 90 kg. The pGH was administered by daily i.m. injection and all pigs were fed one diet. Control animals were pair-fed to the intake noted for pGH-treated pigs between 30 and 60 kg and all pigs were fed ad libitum from 60 to 90 kg. Pigs administered pGH had an improved rate (36%) and efficiency (28%) of gain and an improved protein accretion rate (46%) compared to excipient-treated pigs. Pigs previously treated with pGH continued to exhibit superior (P less than .01) rate and efficiency of gain, and the gain was associated with enhanced protein accretion during the quiescent (postinjection) period compared with excipient counterparts. Administration of pGH between 30 and 60 kg reduced carcass fat and increased carcass protein and water at 90 kg, although fat accretion rate was comparable to that of control pigs. Results indicate that, to varying degrees, the stimulatory effects of pGH on growth performance are sustained following cessation of hormone treatment.  相似文献   

12.
Sixty crossbred pigs (Large White x Landrace) were used in a 2 x 2 x 2 factorial experiment to investigate the effects of gender (intact males vs females) and strain (A vs B) on the response to exogenous porcine growth hormone (pGH) administration (0 [excipient-treated] vs .1 mg pGH.kg live weight-1.d-1). All pigs had ad libitum access to their diet; pGH was administered daily from 60 to 90 kg live weight. All aspects of growth performance and body composition were affected to different degrees by gender and pGH. Strain A pigs had a higher capacity for protein accretion, superior growth performance and contained less fat in the eviscerated carcass and empty body compared with Strain B pigs. Within each strain, intact males ate more feed, had a higher rate of protein deposition and exhibited faster and leaner growth than females. Exogenous pGH administration increased average protein deposition and growth rate by 84 and 34%, respectively, and reduced average feed intake, fat deposition rate, feed:gain and carcass fat content by 14, 59, 37 and 33%, respectively. The magnitude of the changes in growth performance, tissue accretion rates and body composition elicited by pGH were independent of strain. However, within each strain the improvement in feed:gain and reduction in carcass fat measurements elicited by pGH were proportionately larger for females than for intact males.  相似文献   

13.
The present study was undertaken to determine the relationship between dose of porcine growth hormone (pGH) and growth performance of pigs. Porcine GH was administered daily for 35 d [buffer-injected control = (C); 10 micrograms/kg body weight (BW) = (L); 30 micrograms/kg BW = (M); 70 micrograms/kg BW = (H)] to barrows (initial wt = 50 kg). Growth rate was significantly increased by pGH (14% for H dose vs C). Feed efficiency was increased in a dose-related manner (L = 7%, M = 10%, H = 17%) by pGH. There was a concurrent change in carcass composition of pGH-treated pigs. The H dose of pGH decreased the percentage of carcass lipid by 25% (P less than .05). Muscle mass was significantly increased in H vs C pigs (31 vs 26 kg). Serum insulin-like growth factor 1 (IGF-1) concentration increased in a manner that was linearly related to the pGH dose (r = .87). No antibodies to pGH were detected in any of the pigs. In summary, these results extend our earlier findings that pGH increases growth performance markedly. Based on the present findings it appears that the maximally effective dose of pGH is greater than 70 micrograms.kg BW-1.d-1 since several indices of the growth-promoting and metabolic effects of pGH (% carcass protein, % carcass lipid and feed efficiency) had not plateaued.  相似文献   

14.
The current study was undertaken to determine the effects of human growth hormone-releasing factor [hpGRF-(1-44)-NH2] on growth performance in pigs and whether this response was comparable to exogenous porcine growth hormone (pGH) treatment. Preliminary studies were conducted to determine if GRF increased plasma GH concentration after iv and im injection and the nature of the dose response. Growth hormone-releasing factor stimulated the release of pGH in a dose-dependent fashion, although the individual responses varied widely among pigs. The results from the im study were used to determine the dose of GRF to use for a 30-d growth trial. Thirty-six Yorkshire-Duroc barrows (initial wt 50 kg) were randomly allotted to one of three experimental groups (C = control, GRF and pGH). Pigs were treated daily with 30 micrograms of GRF/kg body weight by im injection in the neck. Pigs treated with pGH were also given 30 micrograms/kg body weight by im injection. Growth rate was increased 10% by pGH vs C pigs (P less than .05). Growth rate was not affected by GRF; however, hot and chilled carcass weights were increased 5% vs C pigs (P less than .05). On an absolute basis, adipose tissue mass was unaffected by pGH or GRF. Carcass lipid (percent of soft-tissue mass) was decreased 13% by GRF (P less than .05) and 18% by pGH (P less than .05). Muscle mass was significantly increased by pGH but not by GRF. There was a trend for feed efficiency to be improved by GRF; however, this was not different from control pigs. In contrast, pGH increased feed efficiency 19% vs control pigs (P less than .05). Chronic administration of GRF increased anterior pituitary weight but did not affect pituitary GH content or concentration. When blood was taken 3 h post-injection, both GRF- and pGH-treated pigs had lower blood-urea nitrogen concentrations. Serum glucose was significantly elevated by both GRF and pGH treatment. This was associated with an elevation in serum insulin. These results indicate that increasing the GH concentration in blood by either exogenous GH or GRF enhances growth performance. The effects of pGH were more marked than for GRF. Further studies are needed to determine the optimal dose of GRF to administer in growth trials and the appropriate pattern of GRF administration in order to determine whether GRF will enhance pig growth performance to the extent that exogenous pGH does.  相似文献   

15.
Sixty-six intact male pigs were used to investigate the relationships between exogenous porcine growth (pGH) administration (0, excipient-treated, and .09 mg recombinant pGH.kg-1.d-1) and dietary protein content (8.3, 11.4, 14.5, 17.6, 20.7 and 23.8%) on protein and lipid accretion rates over the live weight range of 30 to 60 kg. Feed intakes were restricted (1.84 kg.pig-1.d-1) and pGH was administered daily by i.m. injection. Rate of protein deposition increased with increasing dietary protein up to 17.6 and 20.7%, respectively, for control and pGH-treated pigs; both growth and protein deposition were enhanced by pGH on the four higher protein diets but remained unaffected by pGH administration to pigs given the two lowest protein diets. Plasma IGF-I concentration was elevated by pGH administration in pigs given the four higher protein diets but unaffected by pGH with the two lowest protein diets. Rate of fat deposition was depressed on all dietary protein treatments by pGH administration; carcass fat content of control and pGH-treated pigs declined with each increase in dietary protein up to 17.6 and 23.8%, respectively. The results demonstrate that pGH acts independently on protein and lipid metabolism.  相似文献   

16.
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17.
Trace mineral status was evaluated in a 2 x 3 factorial treatment array with a total of 34 barrows growing from 25 to 55 kg live weight. Treatments included three levels of feed intake (100, 80 and 60% of ad libitum intake) and exogenous pituitary growth hormone (pGH) therapy (0 and 100 micrograms/kg BW daily). Blood was collected prior to slaughter for the determination of hematocrit and serum trace metal concentrations; tissues (liver, heart, kidney, bone and muscle) were obtained when pigs were slaughtered at 55 kg. Hematocrits and serum Fe were lower in pGH-treated pigs than in controls at all levels of feed intake. Serum Cu was increased by feed restriction but was not altered by pGH therapy. The concentration of serum Zn was not affected by either treatment. Concentrations of hepatic Fe and Cu were lower in pGH-treated pigs than in controls but were higher in feed-restricted pigs than in ad libitum-fed pigs. However, the total amounts of hepatic Fe and Cu were similar in pGH-treated pigs to concentrations in controls. The concentration of hepatic Zn was not influenced by either pGH treatment or feed intake. Femur weights were marginally greater in pGH-treated pigs, probably due to elevated water content. Iron concentration in bone was higher in pGH-treated pigs than in control pigs, whereas Ca, Cu and Zn were not influenced by pGH treatment or feed restriction. Feed intake and pGH treatment did not influence the concentrations of Fe, Cu or Zn in muscle. These findings indicate that pGH therapy affects the metabolism of Fe but has little impact on the overall composition of body ash.  相似文献   

18.
Ten pigs with an average initial live weight of 65 kg were used to investigate the effects of daily exogenous porcine pituitary growth hormone (pGH; .1 mg.kg-1.d-1) for a 13-d period on N retention and whole-body protein turnover. Feed intake was restricted to both the control (treated with excipient) and pGH-treated groups to ensure that animals in each group consumed equal amounts. Whole-body protein turnover was estimated from the excretion of 15N in urinary urea and ammonia after a single oral dose of [15N]glycine. Nitrogen balance and whole-body N flux were increased by 35 to 40% with pGH treatment (P less than .001). Protein synthesis and breakdown were increased by 56 and 59% (P less than .001), respectively, in pGH-treated pigs relative to controls. These higher rates of protein turnover seemed to lower slightly the efficiency of the metabolic process for protein deposition. However, the absolute increment in protein synthesis rate was greater than that for breakdown, leading to the increased net N retention. Thus, pGH treatment improved the utilization of dietary amino acids for protein deposition.  相似文献   

19.
In the present study, pigs were treated daily for 7 days with exogenous porcine growth hormone (pGH; 70 micrograms/kg BW) in order to determine whether pGH induced insulin and insulin-like growth factor 1 (1GF-1) resistance in pig adipose tissue. In the first experiment, pituitary-derived pGH (ppGH) decreased basal and insulin-stimulated lipogenesis by 50%. Insulin sensitivity decreased more than 90% as the result of pGH treatment. Sensitivity and responsiveness to IGF-1 were decreased 50% by ppGH. In a second experiment, pigs were treated daily (70 micrograms/kg BW) with exogenous pituitary pGH (ppGH) or recombinant pGH (rpGH) for 7 days in order to determine if the effects of pGH were intrinsic properties of the hormone. Both rpGH and ppGH caused similar decreases in basal rates of lipogenesis, insulin- and IGF-1-stimulated lipogenesis, and insulin and IGF-1 responsiveness in pig adipose tissue. In summary, the decrease in adipose tissue growth of pigs treated chronically with pGH is due in large part to the suppression of fatty acid synthesis and a decrease in the ability of insulin to stimulate lipid synthesis in pig adipocytes. These responses are intrinsic properties of pGH since the effects of rpGH mimicked those of ppGH. The role and importance of a decrease in IGF-1 responsiveness remains to be resolved.  相似文献   

20.
The acute and chronic effects of porcine growth hormone (pGH) administration on glucose homeostasis of pigs were investigated in the present study. Twelve Yorkshire barrows (average BW = 65 kg) fitted with femoral artery catheters were allotted to three groups. Pigs received acute, intra-arterial injections of either pituitary pGH, a recombinantly derived pGH analog (ppGH or rpGH, 100 micrograms/kg BW) or saline. Acute injection of pGH did not affect fasting plasma glucose or insulin status. Pigs then were treated daily by i.m. injection for 24 d with 70 micrograms ppGH/kg BW. Serum glucose and insulin concentrations during the fed and fasted states were higher in pGH-treated than in control pigs. On d 25, an acute intra-arterial injection of ppGH (100 micrograms/kg BW) elicited increases in plasma glucose and insulin in pigs chronically treated with pGH. The area circumscribed by the glucose and insulin response curves 5 min to 7 h postinjection was 40% (P less than .005) and 177% (P less than .001), respectively, higher in ppGH-treated than in control pigs. These data indicate that pGH does increase plasma glucose and insulin in the fed and fasted states; however, this response is only observed after chronic pGH administration. In addition, pGH is capable of increasing plasma glucose and insulin acutely in the pig. This effect, however, only is observed in pigs treated chronically with pGH. The mechanisms by which pGH elicit these effects on glucose homeostasis are not known.  相似文献   

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