Expression of a recombinant crustacean hyperglycemic hormone of the kuruma prawn Penaeus japonicus in methylotrophic yeast Pichia pastoris

ABSTRACT:   Until now, six crustacean hyperglycemic hormones (CHH) designated Pej-SGP-I, -II, -III, -V, -VI and -VII have been characterized in the kuruma prawn Penaeus japonicus . All CHH consist of 72 amino acid residues and have an amidated carboxyl (C)-terminus. In the present study, we expressed Pej-SGP-III in methylotrophic yeast Pichia pastoris in order to obtain a large quantity of recombinant CHH possessing biological activity. A cDNA encoding Pej-SGP-III that had been previously cloned was processed by polymerase chain reaction (PCR) and the resulting product was ligated into an expression vector. Pichia pastoris was transformed with this vector after which a recombinant Pej-SGP-III was expressed having an additional amino acid residue (glycine) at the C-terminus (rPej-SGP-III-Gly), a form considered to be a putative precursor of this hormone. rPej-SGP-III-Gly secreted into the culture medium was purified by reversed-phase HPLC, and amidated using a peptidylglycine alpha-amidating enzyme. The amidated rPej-SGP-III (rPej-SGP-III) showed hyperglycemic activity in in vivo bioassay almost comparable to that of the natural Pej-SGP-III. rPej-SGP-III thus obtained will be a useful tool not only for its physiological study but also for the determination of its 3-D structure.     

糯米糍荔枝果实内源激素与落果的关系

以糯米糍荔枝生理落果期的田间调查和胚胎发育研究为基础,探讨了内源激素与生理落果的关系.结果表明,糯米糍生理落果高峰的主要原因是:第1次因授粉受精不良导致的生长促进类激素尤其是CTK水平过低;第2次与胚乳为胚的发育所利用而消失、IAA和GAs急剧降低有关;第3次与胚的败育造成CTK、GAs和IAA的全面下降有关;第4次发生在假种皮(果肉)迅速生长期,与种皮和假种皮ABA升至高水平有关.此外,还探讨了ABA /(IAA+GAs+CTK)与生理落果的相关性,以及胚囊、珠被和假种皮等果实内部组分内源激素的变化特点.     

高坡猪GH基因不同基因型对其部分生产性能的影响

采用PCR-RFLP-ApaⅠ方法检测高坡猪GH基因-119~ 486 bp片段的多态性,探讨了不同基因型对其部分生产性能的影响。结果表明:产生2个等位基因A(449 bp)和B(316 133 bp),其频率分别为0.54和0.46;3种基因型:AA、AB和BB,其频率分别为0.26、0.56和0.18;除6月龄腹围和背膘厚外,其余指标值均以BB基因型最高,AA基因型的6月龄腹围和瘦肉率与BB基因型相比差异显著(P<0.05),其他指标在各基因型间差异均不显著(P>0.05)。表明,A可能是小型猪的有利等位基因,B则可能是大型猪的有利等位基因。     

Study on the Tissue Culture of Coleus blumei

This paper studied on the way of Coleus blumei, the leaf was chosen as explants and was inoculated on 1/2 MS medium with different combinations of 6-benzyladenine （6-BA）, α-naphthaleneacetic acid （NAA） and indole-3-butyric acid （IBA）. The optimal conditions of callus induction from explants were achieved on the medium containing 6-BA （2.0 mg· L^-1） and NAA （1.0 mg· L^-1）. Shoot tips were induced on the medium containing 6-BA （4.0 mg·L^-1） and NAA （0.5 mg·L^-1）. The same media conditions were found suitable for shoot multiplication, we multiplied shoots rooted best on 1/2 MS medium supplemented with IBA （0.1 mg·L^-1）.     

提高牦母牛繁殖性能的研究

采用不同剂量生殖激素处理3.5岁的青年牦母牛,结果发情率提高5.84和23.26个百分点,繁殖率分别提高34.69和41.21个百分点。采用不同剂量生殖激素处理和怀孕牦母牛围产期补饲的措施,促进产后成年牦母牛发情配种,结果发情率分别提高42.42,54.67和37.51个百分点,繁殖率提高43.93,48.48和20.10个百分点。生殖激素处理和围产期补饲对提高牦牛繁殖性能有较大的促进作用,高剂量生殖激素的促进效果较明显。     

中间锦鸡儿硬枝扦插试验

以中间锦鸡儿为材料进行了硬枝扦插试验,在珍珠岩和粗沙2种不同的基质上进行扦插,分别采用不同浓度的吲哚丁酸(IBA)、奈乙酸(NAA)、ABT1#、ABT2#生根粉进行处理。试验表明:粗沙硬枝扦插较为理想,平均生根率较珍珠岩提高14.1%;不同激素种类对中间锦鸡儿硬枝插穗生根情况影响极显著,珍珠岩基质上扦插,以ABT1#100 ppm和200 ppm生根效果最佳;粗沙基质上扦插,处理浓度以IBA 1000 ppm、ABT1#500 ppm、ABT2#500 ppm对中间锦鸡儿硬枝扦插生根效果最佳。     

4种外源激素处理对羊草种子萌发和幼苗生长的影响

研究了100~600μg/g赤霉素(GA3)和吲哚乙酸(IAA)、25~100μg/g 6-苄氨基嘌呤(6-BA)和脱落酸(ABA)对羊草种子萌发和幼苗生长的影响。结果表明,100~300μg/g GA3和IAA以及25μg/g 6-BA对羊草种子的萌发最终表现为促进作用,总体效果为GA3>IAA>6-BA。当GA3和IAA浓度大于400μg/g或6-BA浓度大于50μg/g时对羊草种子的最终发芽率均表现为不同程度的抑制;25~100μg/g的外源激素ABA对羊草种子萌发没有抑制作用。GA3处理均能显著促进羊草幼苗的伸长,IAA和6-BA则表现为不同程度的抑制作用;GA3、IAA和6-BA 3种促进激素对羊草的根长和根冠比均表现为抑制作用;25~100μg/g ABA对羊草苗长、根长以及根冠比没有显著影响。     

保幼激素类似物对红火蚁的作用研究进展

我国发现红火蚁已有4年,但防治技术、产品依然匮乏。为了改善、提高我国红火蚁的防治技术水平,本文就保幼激素类似物(JHA)对红火蚁的作用表现和防治效果进行了总结分析。JHA可以造成蚁后卵巢萎缩、产卵量减少,导致发育畸形和蚁群等级比例失调,如工蚁数量减少、生殖型幼虫比例以及处女蚁后数量增加等,并最终导致整个蚁群死亡。JHA杀虫剂活性高、低残毒、对环境污染小,在田间防治红火蚁效果彻底,并可有效地防止防治区红火蚁种群的再入侵（re invasion）,但JHA毒饵也存在着防治效果缓慢等问题。本文就克服此类问题作了初步的探讨。     

Effects of astragaloside on the levels of sex hormone and oxidative stress injury in rats with polycystic ovary syndrome

AIM To investigate the effects of astragaloside on the levels of sex hormone and oxidative stress in rats with polycystic ovary syndrome （PCOS）. METHODS Female SD rats （n=60） were randomly divided into normal control group, model group, Diane-35 （0.339 2 mg/kg） group, low dose astragaloside （12.5 mg/kg） group and high dose astragaloside （50 mg/kg） group, with 12 rats in each group. The PCOS model was induced by letrozole （1 mg/kg）, which was administered by gavage once a day for 3 weeks. After administration, the estrus cycle of the rats was observed by vaginal smear, and the ovarian index was calculated. HE staining was used to observe the histopathological changes of the ovaries. Serum levels of the sex hormones testosterone （T）, estradiol （E₂）, luteinizing hormone （LH） and follicle-stimulating hormone （FSH） were measured by ELISA. The levels of superoxide dismutase （SOD）, malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） in serum and ovarian tissue were detected by colorimetry, and the protein levels of steroidogenetic acute regulatory protein （StAR） and apoptosis-related proteins cleaved caspase-3, Bax and Bcl-2 in ovarian tissue were detected by Western blot. RESULT Compared with control group, the oestrous cycle of the rats in model group was disorder, and the ovarian index was increased, ovary was polycystic. The serum levels of T, LH and MDAwere significantly increased （P<0.05）, while the contents of E2, FSH and the activities of GSH-Px and SOD were significantly decreased （P<0.05）. The levels of MDA, StAR, cleaved caspase-3 and Bax proteins in ovarian tissue were significantly up-regulated （P<0.05）. GSH-Px and SOD activities and Bcl-2 protein levels were significantly down-regulated （P<0.05）. CONCLUSION Astragalosideeffectively balances the levels of sex hormone in PCOS rats and relieves the oxidative stress injury, the mechanism may be related to the inhibition of StAR expression.