凡纳滨对虾热休克蛋白70的原核高效表达

将凡纳滨对虾（Litopenaeus vannamei）热休克蛋白70基因（heat shock protein 70，HSP70）克隆到原核表达载体pET-3c中，经酶切和DNA测序鉴定后，将重组质粒转入表达宿主BL21（DE3），在不同温度、时间下经IPTG诱导表达。收集菌液，进行SDS-PAGE和Western blot检测，并用软件Bandscan分析蛋白表达水平。结果表明，成功构建含凡纳滨对虾HSP70基因的重组表达载体pET-3c／HSP70，表达的目标蛋白相对分子量约为72kD，并能与小鼠抗人HSP70抗体特异性结合。37℃诱导5h目标蛋白表达量最高；但25℃诱导目标蛋白的可溶性比例达80％，目标蛋白占全菌总蛋白70％以上，均较37℃为高。     

Effect of soybean-meal-based diets on the growth and survival rate of the Indian major carp, Cirrhinus mrigala (Ham.)

The effect of varying dietary levels of defatted soybean meal on the growth and survival of mrigal, Cirrhinus mrigala (Hamilton) was investigated. In a feeding trial of 90 days, three experimental diets containing soybean meal at 200, 300 and 400 g kg^?1 level of incorporation were fed to quadruplicate groups of 10 fish each. The conventional feed used in India, consisting of a mixture of groundnut oil cake and rice bran in 1 : 1 ratio served as the control. Best growth in terms of percentage weight gain, specific growth rate, protein efficiency ratio (PER), feed conversion ratio and survival rate was obtained for the test diet with 354 g kg^?1 crude protein and with 400 g kg^?1 soybean meal inclusion level. However, no statistical significant difference was observed between the three soybean‐based diets, except for PER and survival rate. Soybean meal is an easily available, acceptable and cost‐effective protein source in formulated feeds for Indian major carps. The results of the present study indicate that a diet of 350 g kg^?1 overall protein with soybean meal included at 400 g kg^?1 can elicit good growth response and survival in mrigal.     

Dietary arginine requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

Dietary arginine requirement of fingerling Indian major carp, Cirrhinus mrigala (4.20 ± 0.05 cm; 0.60 ± 0.02 g) was determined by conducting a 8‐week feeding trial with casein–gelatine‐based diets (400 g kg^?1 crude protein; 17.90 kJ g^?1, gross energy), containing crystalline amino acids with graded levels of l ‐arginine (10, 12.5, 15, 17.5, 20 and 22.5 g kg^?1, dry diet). Fish were randomly stocked, in triplicate groups, in 55‐L indoor polyvinyl flow through circular tanks and fed experimental diets at 5% of their body weight divided into two feedings at 08.00 and 16.00 hours. Live weight gain (321%) and feed conversion ratio (FCR 1.40) were significantly (P < 0.05) higher in fish fed diet containing 17.5 g kg^?1dietary arginine compared with other diets. Second‐degree polynomial regression analysis of live weight gain, FCR and protein efficiency ratio data indicated requirements for dietary arginine at 18.7, 18.4 and 18.3 g kg^?1 of the dry diet, respectively. Maximum carcass protein, and minimum moisture and fat contents were noticed at the requirement level. Carcass ash content remained insignificantly different among the treatments except at 17.5 g kg^?1 dietary arginine showing significantly higher ash content. Based on the above results, it is recommended that the diet for fingerling C. mrigala should contain arginine at 18.4 g kg^?1, dry diet, corresponding to 46 g kg^?1 dietary protein for optimum growth and efficient feed utilization.     

Dietary threonine requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

Indian major carp fingerling, Cirrhinus mrigala (3.85±0.75 cm, 0.52±0.21 g), were fed isonitrogenous and isocaloric diets (40% crude protein, 4.28 kcal g^?1, gross energy) containing casein, gelatin and crystalline amino acids with graded levels of l ‐threonine (1.00, 1.25, 1.50, 1.75, 2.00 and 2.25 g 100 g^?1, dry diet) to determine the dietary threonine requirement. The feeding trial was conducted in triplicate for 8 weeks. Diets were fed twice a day at 08:00 and 16:00 hours at 5% body weight day^?1. The ration size and feeding schedule were worked out before the start of the feeding trial. Highest weight gain (304%) and best feed conversion ratio (1.43) were evident in fish fed diet containing 1.75% dietary threonine. Second‐degree polynomial regression analysis of weight gain, feed conversion ratio and protein efficiency ratio data indicated the dietary threonine requirement to be at 1.84%, 1.81% and 1.78%, respectively, corresponding to 4.60%, 4.52% and 4.45% of dietary protein. Minimum carcass moisture, fat and maximum carcass protein were evident in fish fed 1.75% threonine level. However, ash content did not affect body composition, except the 1.00% threonine level, which showed a significantly higher ash content value. Based on the above results, it is recommended that the diet for C. mrigala should contain threonine at 1.80 g 100 g^?1 dry diet, corresponding to 4.50 g 100 g^?1 dietary protein for optimum growth and efficient feed utilization.     

脊尾白虾热休克蛋白HSP70基因的克隆及其表达分析

克隆了脊尾白虾热休克蛋白基因全长cDNA,并进行了序列分析。该基因由2 250 bp的碱基组成,开放阅读框长1 959 bp,编码由652个氨基酸组成的蛋白,基因两翼分别存在83 bp(5′端)和208 bp(3′端)的非翻译区,将该基因命名为Ec-HSP70。与其它物种HSP70s氨基酸序列进行同源性比较发现,与甲壳动物的同源性都在90%以上,表明该蛋白属于热休克蛋白HSP70家族。聚类分析表明,脊尾白虾热休克蛋白氨基酸序列与中国明对虾和凡纳滨对虾紧密聚为一支,之后聚类顺序依次为斑节对虾、日本囊对虾、刀额新对虾等。通过荧光定量RT-PCR对该基因在肝胰腺、肌肉的表达分析表明,温度、pH和氨氮胁迫都可以引起该基因的高表达,而且肝胰腺中的高表达时间相对肌肉中出现的较早。试验结果表明,温度、pH和氨氮胁迫对脊尾白虾HSP70基因表达有一定的诱导效果,但胁迫的时间过长则抑制其表达,肝胰腺对胁迫比肌肉较敏感。     

Dietary histidine requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

An 8‐week growth trial was conducted to determine the dietary histidine requirement of the Indian major carp, Cirrhinus mrigala fingerling (length 4.22 ± 0.45 cm; weight 0.61 ± 0.08 g; n = 40). Isonitrogenous (400 g kg^?1 crude protein) and isoenergetic (17.90 kJ g^?1 gross energy) diets with graded levels of l ‐histidine (2.5, 5.0, 7.5, 10.0, 12.5 and 15.0 g kg^?1 dry diet) were formulated using casein and gelatin as a source of intact protein, supplemented with l ‐crystalline amino acids. Twenty fish were randomly stocked in 70‐L indoor polyvinyl circular fish tank (water volume 55‐L, water exchange rate 1–1.5 L min^?1) and fed experimental diets at the rate of 5% of their body weight/day divided over two feedings at 08:00 and 16:00 h. Maximum live weight gain (295%), best feed conversion ratio (FCR) (1.48) and protein efficiency ratio (PER) (1.69) occurred at 7.5 g kg^?1 of dietary histidine level. When live weight gain, FCR and PER data were analysed using second‐degree polynomial regression, the break points indicated histidine requirements at 9.4, 8.6 and 8.5 g kg^?1 of dry diet respectively. Significantly (P < 0.05) higher whole body protein and low moisture values were recorded at 7.5 g kg^?1 histidine level. Body fat increased significantly (P < 0.05) with increasing histidine levels. However, at 7.5 and 10 g kg^?1 histidine diets body fat did not differ (P > 0.05) to each other. Ash content of fish fed diets containing various levels of histidine did not differ except at 2.5 and 5.0 g kg^?1 inclusion levels where significantly (P < 0.05) higher ash was recorded. Protein deposition was also found to be significantly (P < 0.05) higher in the 7.5 g kg^?1 histidine diet. Based on the polynomial regression analysis of FCR and PER data, it is recommended that the diet for fingerling C. mrigala should contain histidine at 8.5 g kg^?1 of dry diet, corresponding to 21.25 g kg^?1 of dietary protein for optimum growth and efficient utilization of feed.     

Dietary tryptophan requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

An 8‐week feeding trial was conducted to evaluate the effects of dietary tryptophan concentration on weight gain and feed efficiencies of fingerling Indian major carp, Cirrhinus mrigala. Six isonitrogenous (40% crude protein) and isocaloric (17.90 kJ g^?1) amino acid test diets containing casein, gelatin and l ‐crystalline amino acids with graded levels of l ‐tryptophan (0.06, 0.16, 0.26, 0.36, 0.46 and 0.56 g 100 g^?1 dry diet) were formulated. Fish (4.25±0.30 cm, 0.62±0.02 g) were randomly stocked in triplicate groups in 70 L (water volume 55 L) flow‐through (1–1.5 L min^?1) indoor circular tanks and fed experimental diets at 5% of their body weight/day in two feedings at 08:00 and 16:00 hours. Maximum live weight gain (277%), lowest feed conversion ratio (FCR) (1.50) and highest protein efficiency ratio (PER) (1.66) were measured at 0.36% dietary tryptophan. The relationship between dietary tryptophan levels and weight gain, FCR and PER data were described using second‐degree polynomial regression analysis indicating the tryptophan requirement at 0.42, 0.39 and 0.38 g 100 g^?1 of dry diet respectively. Whole body moisture decreased with increasing tryptophan up to 0.36%. Significantly (P<0.05) higher protein content was evident in fish fed diet containing 0.36% tryptophan. Body fat increased significantly (P<0.05) in fish fed with different tryptophan concentrations except those fed 0.36% tryptophan where a significantly lower fat content was noted. Significantly (P<0.05) higher ash content was reported at 0.06% and 0.16% tryptophan levels. Survival was 100% in fish fed all the diets except those fed 0.06% tryptophan. Based on the results, diets for fingerling C. mrigala should contain tryptophan at 0.38 g 100 g^?1 dry diet, corresponding to 0.95 g 100 g^?1 dietary protein for optimum growth and efficient feed utilization.     

Dietary methionine requirement of Indian major carp fry,Cirrhinus mrigala (Hamilton) based on growth,feed conversion and nitrogen retention efficiency

This study was aimed at quantifying methionine requirement of Indian major carp fry, Cirrhinus mrigala (2.2 ± 0.2 cm; 0.19 ± 0.02 g) by conducting a 12‐week feeding trial. Casein–gelatine‐based isonitrogenous (40 g 100 g^?1 crude protein) and isoenergetic (15.42 kJ g^?1 DE) amino acid test diets were prepared to contain six levels of l ‐methionine (1.1, 1.3, 1.5, 1.7, 1.9 and 2.1 g 100 g^?1 dry diet) at a fixed level of cysteine (0.85 g 100 g^?1 dry diet) and fed to apparent satiation thrice daily to triplicate groups of fish. When absolute weight gain (g per fish), feed conversion ratio, protein deposition (g per fish) and nitrogen retention efficiency data were subjected to broken‐line and second‐degree polynomial regression analysis, 95% of the plateau of above parameters was achieved at dietary methionine concentrations between 1.60 and 1.69 g 100 g^?1 dry diet or 0.10 to 0.11 g methionine kJ^?1 DE, corresponding to 4.1–4.22 g 100 g^?1 protein or 0.44–0.47 g methionine kJ^?1 DE. Based on these results, dietary methionine requirement of fry C. mrigala is recommended 1.60–1.69 g 100 g^?1 diet or 0.10–0.11 g methionine kJ^?1 DE.     

Effect of ration size on growth, conversion efficiency and body composition of fingerling mrigal, Cirrhinus mrigala (Hamilton)

Optimum ration size of Indian major carp, Cirrhinus mrigala fingerlings was determined by feeding purified diet (40% crude protein (CP); 3.61 kcal g^?1) at five ration sizes (2%, 4%, 6%, 8% and 10% of body weight per day) for 6 weeks. Feeding trial was conducted in triplicate. Fishes were randomly stocked at the rate of 20 fish per trough fitted with water flow‐through system. The best feed conversion ratio (FCR), specific growth rate (%) and protein efficiency ratio (PER) were evident at the ration size of 4–6% body weight. Second‐degree polynomial regression analysis of the FCR, PER, and protein and energy retention data indicated the breakpoints at ration size of 5.16%, 5.24%, 5.52% and 5.42% body weight per day. Carcass composition of fish fed different ration sizes varied significantly. Maximum carcass protein and minimum moisture content were noticed at 4% and 6% ration levels. A linear increase in fat content was evident with increasing ration levels up to 6% body weight. Ash content remained insignificantly different among various ration levels except at 2%, showing the significantly highest value. Water temperature, dissolved oxygen, free carbon dioxide, pH and total alkalinity were recorded regularly during the length of the experiment. No mortality was observed during the feeding trial. Based on the above results, it is recommended that feeding in the range of 5–5.5% body weight per day corresponding to 20 g protein and 181 kcal energy to 22 g protein and 199 kcal energy per kg of the diet per day is optimum for the growth and efficient feed utilization of C. mrigala.     

热应激对剑尾鱼HSP70家族两成员基因表达的影响

采用RT-PCR方法扩增实验动物剑尾鱼（Xiphopohorus helleri）纯系RR-B的HSP70家族两个成员的eDNA片段，并将其克隆到PMD18-T载体中测序，将测序结果与GenBank中的核苷酸序列和推导的氨基酸序列进行同源性比较。同时利用RT-PCR半定量方法研究热应激时两个成员在剑尾鱼组织中的表达。通过克隆获得了剑尾鱼的HSP70家族两个新成员的cDNA片段，两序列均包含HSP70家族的特征性签名序列和热休克蛋白的定位序列；通过对克隆片段与已发表的青锵（Oryzias latipes）等鱼类HSP70的核苷酸序列和其编码的氨基酸序列同源性比较，发现核苷酸序列同源性较高，成员一和成员二分别为85％-89％和82％-99％；氨基酸序列同源性更高，成员一和成员二分别为97％-99％和93％-99％。一般条件下，两成员在剑尾鱼肝脏、脾脏、肾脏和心脏中不表达，但热应激能刺激成员一在剑尾鱼脾脏中表达，成员二在肝脏、脾脏、肾脏和心脏中强烈表达，并可能在参与热应激保护方面起更大作用。     

合浦珠母贝热休克蛋白hsp70基因的克隆与表达分析

采用同源克隆和RT-PCR技术对合浦珠母贝(Pinctada fucata)热休克蛋白hsp70基因进行了克隆和表达分析。获得cDNA全长序列2 365 bp,其中3’非编码区域(UTR)为318 bp,5’UTR为88 bp,开放阅读框(ORF)为1 959 bp,编码652个氨基酸,分子量约为71.39 kD,理论等电点为5.22,并含有3个HSP70家族的签名序列IDLGTTYS、DLGGGTFD和EEVD。同源性分析表明,合浦珠母贝HSP70的氨基酸序列与太平洋牡蛎(Crassostrea gigas)等双壳贝类的相似性高达86%以上,基于氨基酸序列的聚类分析表明,合浦珠母贝与牡蛎属种类亲缘关系最近。高温、高盐刺激后,半定量RT-PCR检测发现hsp70基因的表达明显增加,高温刺激的表达量高于高盐刺激,高温刺激组不同组织的表达量由大到小依次为鳃、消化腺、外套膜、肌肉、性腺,高盐刺激组不同组织的表达量由大到小依次为鳃、外套膜、肌肉、消化腺、性腺,表明HSP70参与了机体对刺激的应答过程。该基因的克隆为进一步深入研究合浦珠母贝的抗逆机理及其遗传改良奠定了重要基础。     

Acute toxicity of ammonia and its sub-lethal effects on selected haematological and enzymatic parameters of mrigal, Cirrhinus mrigala (Hamilton)

A comprehensive acute toxicity trial was conducted using a static water system to study the toxic effect of ammonia on haematology and enzyme profiles of Cirrhinus mrigala H. The LC₅₀ of total ammonia‐nitrogen (TAN) was 11.8 mg L^?1 TAN (1.029 mg L^?1 NH₃‐N). The sub‐lethal test revealed that with increasing concentration of TAN, the total erythrocyte counts were reduced in lower concentrations (1–4 mg L^?1 TAN) followed by higher levels in fish exposed to higher concentrations (8–16 mg L^?1 TAN). In contrast, the total leucocyte counts were opposite. With increasing concentration of TAN, haemoglobin and serum protein content were reduced, whereas the blood glucose level increased. As the concentration of ammonia increased, there was a reduction in acetylecholinesterase activity in the brain and liver; alkaline phosphatase activity in the serum, brain and gill; and acid phosphatase (ACP) activity in the gill. The activity of lactate dehydrogenase in the gill, liver, kidney and brain increased with increased concentration of ammonia. In addition, activities of ACP in the serum and brain, alanine aminotransferase in the serum, brain and gill, and aspartate aminotransferase in the serum, brain and gill were increased.     

Reproduction phase-related variations in the GnRH immunoreactive fibers in the pineal of the Indian major carp Cirrhinus mrigala (Ham.)

We studied GnRH immunoreactivity in the pineal gland of the Indian major carp Cirrhinus mrigala during different phases of reproductive cycle. In the resting phase (December–January), GnRH immunoreactive (-ir) fibers were organized as paired fascicles above the posterior commissure that ascend in the stalk and distribute widely in the pineal gland. The GnRH-ir fiber density significantly declined (P<0.001) during the preparatory phase (February–April) and the fibers disappeared thereafter. While no GnRH fibers were seen during the prespawning (May–June) and spawning (July–August), isolated GnRH-ir fibers reappeared in the postspawning phase. Since no GnRH cell bodies were detected in the pineal, these GnRH-ir fibers seem to be of central origin. The results reveal a distinct reciprocal relationship between the GnRH immunoreactivity in the pineal and the status of the ovarian maturity; the fibers appeared in the pineal only during the period of ovarian quiescence. While the functional significance of these cyclic changes in GnRH is yet to be determined, we suggest that the decapeptide may serve as a transmitter of central origin that modulates the activity of the pineal gland.     

Specific immune response in mucosal and systemic compartments of Cirrhinus mrigala vaccinated against Edwardsiella tarda: In vivo kinetics using different antigen delivery routes

Mucosal immune barriers confer protection against invading fish pathogens. Here, we conducted an experiment for 60 days to assess the mucosal and systemic immune response in Mrigal (Cirrhinus mrigala), an Indian major carp. Fish were immunized with inactivated Edwardsiella tarda by four different routes, namely, oral, immersion, injection, and anal intubation. An indirect enzyme‐linked immunosorbent assay (ELISA) was used to measure the specific immune response (antibody) in serum and mucus (collected from skin, gill, and gut) of the fish on 0, 15, 30, 45, and 60 days postimmunization. For specific immune response in the serum, significantly higher (p < 0.05) optical density (OD) values were obtained in the anal group (0.52 ± 0.03) and in the oral group (0.48 ± 0.03). In the skin mucus, significantly higher OD values were obtained in the oral group (0.48 ± 0.04) and immersion group (0.32 ± 0.03). In the gill mucus, significantly higher OD values were obtained in the oral group (0.82 ± 0.08) and the immersion group (0.73 ± 0.03). In the gut mucus, significantly higher OD values were obtained in the immersion group (0.080 ± 0.007) compared to the rest of the treatments. Fish from all the groups were challenged with LD₅₀ dose of E. tarda at the end of the experiment. We conclude that oral and immersion immunization routes offer better protection of C. mrigala compared to other antigen delivery routes.     

团头鲂 HSP70 cDNA 的克隆、序列分析以及热应激对其 mRNA 表达的影响

采用RT-PCR和cDNA末端快速扩增法(RACE)分离、克隆出团头鲂(Megalobrama amblycephala)肝脏热休克蛋白70(HSP70)基因全长cDNA序列,共2 224 bp,包括135 bp5'非翻译区、1 932 bp阅读框以及157 bp 3'非翻泽区[不包括Poly(A)].阅读框共编码643个氨基酸,分子量计算值为70.53 kD,理论等电点为5.25.该HSP70基因在翻译区不含内含子,符合诱导型HSP70的特征.团头鲂HSP70氨基酸序列中含有HSP70家族的3个签名序列以及细胞质特异性调控基序EEVD等.同源性分析表明,团头鲂HSP70氨基酸序列与斑马鱼等脊椎动物的相似性高达84.0%以上,与无脊椎动物果蝇的相似性也高达73.4%.生物信息学分析显示,团头鲂HSPTO基因所编码的蛋白质以亲水性区域为主,具有丰富的B细胞抗原位点,无信号肽,无跨膜区域,为非分泌型蛋白;同时还含有众多的蛋白激酶C磷酸化位点、N-肉豆蔻酰化位点、酪蛋白激酶Ⅱ磷酸化位点和N-糖基化位点,推测其可能在细胞信号转导与调控中发挥重要作用.该蛋白含有2个部分重叠的双向核定位序列,二级结构以α-螺旋和无规卷曲为主,空间结构包括N端ATPase功能域和C端多肽结合功能域.应用实时定量PCR研究高温(34℃)应激对团头鲂HSP70 mRNA表达的影响.结果表明,在热应激过程巾肝脏HSP70 mRNA水平呈先升高后下降的变化趋势,说明该基因的表达受热应激调控,本实验克隆的序列为诱导型HSP70 cDNA.     

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.     

热应激对大正三色锦鲤非特异性免疫指标及HSP70基因表达的影响

为探讨锦鲤的抗逆机理,在实验室条件下研究了持续热应激对大正三色锦鲤非特异性免疫指标及HSP70基因相对表达量的影响。分别于应激前、应激后2、6、10、14、18、22、26 h进行取样测定呼吸爆发、补体蛋白3(C3)、超氧化物歧化酶(SOD)、丙二醛(MDA)以及热休克蛋白70(HSP70)基因的相对表达量。结果发现,热应激后呼吸爆发降低,在应激后2、10、18～26h降低显著(P<0.05);热应激2 h后血清中C3含量略有上升,应激6～26 h过程中与应激前相比均下降,应激14～22 h显著地下降(P<0.05);热应激下血清中SOD有上升的趋势,但差异不显著(P>0.05);热应激下,血清中MDA浓度升高,应激10～18 h上升显著(P<0.05);应激后2、26 h HSP70基因的相对表达量与应激前相比显著性地上升了5.93倍、2倍(P<0.05),应激后6～22 h HSP70基因的相对表达量与应激前水平无显著性差异(P>0.05)。结果表明,热应激影响锦鲤非特性免疫指标,降低锦鲤非特异性免疫力;HSP70的表达受热应激调节,热应激下诱导合成的HSP70对锦鲤起到一定的应激保护作用。     

KK-42对凡纳滨对虾热激蛋白基因表达的诱导作用

为探讨咪唑类物质KK-42可能的作用机制,首次克隆了热激蛋白90(HSP90)部分mRNA序列,研究了HSP90以及HSP70基因的时空表达以及KK-42对其表达的影响。将体长3.5～5.0 cm凡纳滨对虾幼虾随机分为2组,分别用1.95×10^-4 mol/L的KK-42溶液或不含KK-42的溶液浸泡处理1 min,之后在相同条件下常规饲养。结果显示, HSP90和HSP70在凡纳滨对虾大颚器官、眼柄、肌肉和肝胰腺都有表达,其中,肝胰腺中的表达水平较高。实验期间,肝胰腺中两种HSP mRNA水平虽有一定波动,但含量相对较低;KK-42处理可显著诱导肝胰腺HSP的表达,在处理后第1、2、3天,HSP70和HSP90 mRNA含量分别比相应的对照组升高了243.4%、141.3%、410.5%和121.6%、505.5%、481.7%。结果表明,KK-42处理可显著提高凡纳滨对虾肝胰腺HSP90和HSP70基因的转录水平。