豇豆胰蛋白酶抑制剂转基因棉花的获得

利用根癌农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍｔｕｍｅｆａｃｉｅｎｓ）介导法将豇豆胰蛋白酶抑制剂（Ｃｏｗ－ｐｅａＴｒｙｐｓｉｎＩｎｈｉｂｉｔｏｒ,ＣｐＴＩ）基因转移进入棉花（ＧｏｓｙｐｉｕｍｈｉｒｓｕｔｕｍＬ．）。棉花幼苗的下胚轴与携带有ＣｐＴＩ基因和Ｎｐｔ－Ⅱ基因的根癌农杆菌共培养,在选择培养基上诱导出了卡那霉素抗性（Ｋｍｒ）愈伤组织。选择Ｎｐｔ－Ⅱ阳性的胚性愈伤组织在胚状体诱导培养基上进行胚状体诱导,继而在分化培养基上进行植株分化,最终获得了再生棉花植株。经Ｎｐｔ－Ⅱ分析、ＰＣＲ及Ｓｏｕｔｈｅｒｎ检测证明,外源ＣｐＴＩ基因和标记基因（Ｎｐｔ－Ⅱ基因）存在于转化棉株及其后代中。抗棉铃虫（Ｈｅｌｉｏｔｈｉｓａｒｍｉｇｅｒａ）生物活性检测表明转基因植株后代具有明显的抗棉铃虫能力。     

Intracellular and extracellular enzyme activity in soil with reference to elemental cycling

Enzyme activities play an important role for the transformation of elements and compounds in soil and, thus, were extensively analyzed for more than 4 decades. The activity of any enzyme in soil may not only be controlled by active organisms. Substantial parts of ‘extracellular’ enzymes may be stabilized by abiotic soil components maintaining their activity. Methods to discriminate the source of enzyme activity were summarized with emphasis on the approach plotting enzyme activity versus a feature integrating the microbial biomass after the addition of glucose and nitrate. Considering the quotient between enzyme activity and microbial biomass content, protease activity will be discussed with reference to nitrogen transformation in soils.     

碱性蛋白酶水解猪血细胞最优反应条件的研究

研究旨在确立碱性蛋白酶水解猪血细胞的最优反应条件。采用二次旋转正交回归设计,建立了ｐＨ值、温度（Ｔ）、酶—底物浓度比（〔Ｅ〕／〔Ｓ〕）三个因素对水解蛋白收率影响的回归模型。通过分析,确定最优水解条件为：ｐＨ＝８２１,〔Ｅ〕／〔Ｓ〕＝４％,最后,在此最优水解条件下,确定出最佳水解时间ｔ＝９０ｍｉｎ。     

胰蛋白酶对叶蛋白水解作用的研究

以植物叶蛋白浓缩物为材料,用胰蛋白酶在不同条件下对其进行水解。结果表明,胰蛋白酶作用于植物叶蛋白,其最佳水解温度为４０℃;最佳水解ｐＨ值为９;水解程度随酶量和反应时间增加而增加;各种不同种类的植物叶蛋白在相同条件下被胰蛋白酶水解的程度基本一致,证明了植物叶蛋白易被动物体内蛋白酶分解。     

蚕蛹蛋白双酶水解工艺条件研究

蚕蛹脱脂后,用微生物酶进行双酶水解,确定其最佳工艺条件为：３９４２中性蛋白酶反应温度为４０℃,ｐＨ为７０,固液比为２０∶１００,加酶量为２０％,反应时间为２ｈ;１３９８中性蛋白酶反应温度为５０℃,ｐＨ为７５,固液比为２０∶１００,加酶量为２０％,反应时间为１ｈ．在该条件下水解,水解率高达２３７５％,水解液中的１８种氨基酸质量浓度为１５６４ｇ·Ｌ－１．     

维生素B6对史氏鲟幼鱼胰蛋白酶活性的影响

试验研究了饲料中不同水平维生素B6对史氏鲟（Acipenser schrencki）幼鱼胰蛋白酶活性的影响。在温度为（20±0.5）℃、pH值为6.8～7.5的条件下,研究了低蛋白含量（28%）饲料中,添加不同水平的维生素B6（0、15、30、45、60、75mg/kg）对史氏鲟幼鱼[（34.07±3.44）g]胰蛋白酶活性的影响。结果表明,饲料中的维生素B6添加量为45～60mg/kg时,对幼鲟胰蛋白酶的激活作用最大,并可相应提高对饲料蛋白质的消化率。     

Effect of water temperature on the growth performance and digestive enzyme activities of Chinese longsnout catfish (Leiocassis longirostris Günther)

The present study was carried out to investigate the influence of water temperature on the growth performance and digestive enzyme (pepsin, trypsin and lipase) activities of Chinese longsnout catfish. Triplicate groups of Chinese longsnout catfish (35.6±0.48 g, mean±SE) were reared at different water temperatures (20, 24, 28 and 32 °C). The feeding rate (FR), specific growth rate (SGR) and feed efficiency ratio (FER) were significantly affected by water temperatures and regression relationships between water temperature and FI, SGR as well as FER were expressed as FR=−0.016 T ²+0.91 T −10.88 ( n =12, R ²=0.8752), SGR=−0.026 T ²+1.39 T −17.29 ( n =12, R ²=0.7599) and FER=−0.013 T ²+0.70 T −8.43 ( n =12, R ²=0.7272). Based on these, the optimum temperatures for FR, SGR and FER were 27.66, 26.69 and 26.44 °C respectively. The specific activities of digestive enzymes at 24 or 28 °C were significantly higher than that at 20 or 32 °C. In addition, there was a significant linear regression between FR or SGR and specific activities of pepsin and lipase, which indicated that pepsin and lipase played important roles in regulating growth through nutrient digestion in Chinese longsnout catfish.     

不同生长阶段兰州鲇消化酶活性的比较研究

测定了不同生长阶段兰州鲇(Silurus lanzhotaensis)胃、肝胰脏、前肠、中肠、后肠的蛋白酶、淀粉酶和脂肪酶的活性.结果表明,兰州鲇蛋白酶活性大小为:前肠>中肠>后肠>肝胰脏>胃,阶段1[体重(51.9±6.0)g]>阶段2[体重(228.6±16.6)g]>阶段3[体重(448.9±26.5)g],且差异显著;淀粉酶活性:肝胰脏>胃>前肠>中肠>后肠,阶段1>阶段2>阶段3,差算显著;脂肪酶活性大小为:后肠>中肠>前肠>肝胰脏>胃,但各阶段差异不显著.随着年龄的增长,兰州鲇对蛋白质和碳水化合物的消化能力逐渐减弱.     

Synthetic tetramer of a Phytophthora sojae-inducible fragment from soybean GmaSKTI36 promoter improves its pathogen induction activities

Using pathogen-induced promoters to control expression of the functional genes in transgenic plants may greatly increase the chances of boosting disease resistance. However, the number of the inducible promoters is limited. Here, we found that soybean GmaSKTI36 gene is strongly induced upon Phytophthora sojae infection. Functional analysis showed that its promoter could mediate rapid and strong induction of GUS expression upon pathogen infection in both Nicotiana benthamiana leaves and soybean hairy roots. Then, a 122 bp fragment that was critical to the activity was successfully identified by a progressive 5′ deletion analysis. Importantly, we found that a synthetic promoter by tetramerizing this fragment could confer strong P. sojae induction activities. Overall, the results suggested that the GmaSKTI36 promoter, the 122 bp fragment, and the synthetic promoter are potentially useful pathogen-inducible promoters.     

Ontogenic changes in the digestive enzyme patterns and characterization of proteases in Indian major carp Cirrhinus mrigala

Digestive enzymes of Cirrhinus mrigala (Ham.) were studied during ontogenic development. Specific amylase activity was detected in first feeding fish. The enzyme activity decreased up to day‐18 and then it increased with the age of fish to reach the highest level on day‐34. Protease activity was 28.61 ± 8.90 mU mg protein^?1 min^?1 on day‐4 and increased with the age throughout the study period. Trypsin activity was 31.86 ± 1.12 mU mg protein^?1 min^?1 on day‐4. The activity decreased up to day‐10 and from day‐12 onwards increased up to day‐26. Chymotrypsin activity was 14.56 ± 2.74 mU mg protein^?1 min^?1on day‐4 and constantly increased up to day‐26. A significant increase in lipase activity was observed between days‐24 and 34. SDS‐PAGE and substrate SDS‐PAGE showed the diversity of protein (17.4–127.8 kDa) and protease activity bands (16.6–88.8 kDa) during ontogenesis. Soybean trypsin inhibitor, phenyl methyl sulphonyl fluoride, N‐α‐p‐tosyl‐l ‐lysine chloromethylketone and N‐tosyl‐l ‐phenylalanine chloromethylketone inhibited the protease activity up to 79.72–97.21, 65.55–94.83, 45.41–75.31 and 40.78–64.72%, respectively. Inhibition study in substrate SDS‐PAGE revealed the abundance of serine proteases and the presence of isoforms of trypsin and chymotrypsin. Ethylenediamine‐tetraacetate showed 5.56–22.78% inhibition of metal ion‐specific enzyme activity.