分子文库展示技术

分子文库展示技术是一系列广泛应用于多肽、蛋白质及药物筛选和研究蛋白质间相互作用的有效的生物学技术。它将组合成的具有一定长度的随机序列寡核苷酸片段(或cDNA)克隆到特定表达载体中,使其表达产物(多肽片段或蛋白质结构域)以融合蛋白的形式展示在活的噬菌体或细胞表面。根据其蛋白质表达是否依赖于宿主表达系统,分为体内表达展示系统和无细胞展示系统(体外表达展示系统)。就其展示的部位不同又可分为噬菌体展示技术、细胞表面展示技术、核糖体展示技术、mRNA展示技术等。现对各种展示技术的基本原理及相关应用做简要综述。     

我国蜜蜂主要病原检测技术

蜜蜂是重要的经济昆虫。蜜蜂病害威胁蜜蜂产业的发展。蜜蜂病害检测及病原鉴定是病害防治的基础。文章详细介绍我国蜜蜂常见6种主要病害(美洲幼虫腐臭病,欧洲幼虫腐臭病,囊状幼虫病,慢性麻痹病,微孢子虫病,白垩病)的病原快速准确的检测方法。     

合成生物制造2022

本文对2022年《生物工程学报》发表的与合成生物制造相关的综述和研究论文进行了评述,重点讨论了DNA测序、DNA合成、DNA编辑、基因表达调控和数学细胞模型等底层技术,酶的设计、改造和应用技术,化学品生物催化、氨基酸及其衍生物、有机酸、天然化合物、抗生素与活性肽、功能多糖、功能蛋白质等重要产品的生物制造技术,一碳化合物和生物质原料利用技术以及合成微生物组技术,以帮助读者从一个侧面了解合成生物制造相关技术和产业的发展情况。     

史前古人类之间的基因交流及对当今现代人的影响

古DNA实验技术及高通量测序技术的出现和发展,使得直接从古老化石中进行遗传物质的提取及测序成为可能,与古人类相关的基因组学研究因此取得了一系列突破性进展,已灭绝的古老型人类(如:尼安德特人和丹尼索瓦人)与非洲以外现代人之间基因的相互影响已被诸多证据所证实。研究表明,在史前时期,早期现代人向非洲以外地区扩散时,遭遇到了现已灭绝的古老型人类,他们在同一时空内长期共存,并发生了基因交流,有一部分古老型人类基因因此流向了现代人,有些基因一直流传至今,对当今现代人的基因组成产生重大影响;此外,不同古老型人类之间也存在基因交流;而早期现代人也对部分古老型人类的基因组成造成了影响。化石与古DNA信息的证据均表明,史前各种人类之间的基因交流在多个地区发生多次,他们的基因交流共同构建了当今现代人的基因库,并在生理机能、形态和疾病发生率等方面对现代人造成了深远的影响。     

Influence of soil type,crop and air drying on residual carbohydrate content and aggregate stability after treatment with periodate and tetraborate

Summary The relationship between the water stability of microaggregates and the residual carbohydrate content of soil was examined in 15 soils from 7 soil series under various cultivations. The carbohydrate was progressively removed by increasing the time of treatment with 0.02M periodate and 0.1M tetraborate. The resulting decrease in reducing sugar content was significantly correlated with an increased disruption of microaggregates (>45 m) as determined by a turbidimetric method. The most effective treatment removed about 80% of the soil carbohydrate and caused an increase of about 75% in the fraction of microaggregates (<45 m) compared to untreated soil.15–20 percent of the soil carbohydrate was resistant to oxidation by periodate, even after prolonged reaction times and contained a higher relative proportion of glucose, arabinose, and xylose than the oxidised material. Sugars typical of microbial sources, mannose, galactose, rhamnose and fucose, were therefore preferentially oxidised by the periodate treatment.The grassland soils generally had higher carbohydrate contents than the arable soils and initially had a greater degree of aggregation. However, periodate oxidation affected each soil in its own characteristic manner. A significant inverse linear relationship between the degree of disruption and the residual sugar content was found with 13 of the 15 soils. Over the range measured aggregate stability was therefore related to the presence of carbohydrate predominantly from microbial sources.     

Low invasive estrous synchronization protocol for wild animals: an example with melengestrol acetate in brown brocket deer (Mazama gouazoubira)

Deer are sensitive to stressful stimuli by handling and their reproductive physiology could be altered by these procedures, making it necessary to develop less invasive protocols for ART. Melengestrol acetate (MGA), a synthetic progestin administered orally, appears as an alternative for estrous synchronization protocols (ESP), such as reported in cattle. Firstly, we compared two MGA doses (0.5 and 1.0 mg/day/animal), which would have suppression effect in estrous behavior (EB). Eight females were randomly and equally distributed in Group 1 (G1) and Group 2 (G2), which received 0.5 and 1.0 mg/day/animal respectively for 15 days (D1 to D15). Two cloprostenol (CP) applications were performed on D0 and D11. Estrus detection (ED) was performed every day. All females from G1 displayed estrus during treatment period, whereas all females from G2 displayed estrus after treatment, suggesting a suppressive effect of 1.0 mg in the EB. Once the suppressive MGA dose (1.0 mg) was defined, we used this dose for assessing ESP. The same eight females received 1.0 mg/animal for eight days (D-8 to D-1), followed by 0.25 mg of estradiol benzoate on D-8 and 265 μg of CP on D0. Feces for fecal progesterone metabolites (FPM) measurement were collected from D0 until seven days after the last day of estrus. Seven females displayed estrus between 12 and 72 h after CP application, which was followed by a significant increase in FPM levels (except female MG6), suggesting the formation of corpus luteum. After ED, females were placed with a fertile male to assess the fertility of the protocol. Pregnancy was confirmed by ultrasound 30 days after mating in 3/6 individuals. Although the low effectiveness of MGA protocol, it should be considered as a promising alternative in deer ESP since this protocol has less stressful effect on the animal during reproductive management when compared to other ESP.     

Native seeds incorporated into activated carbon pods applied concurrently with indaziflam: a new strategy for restoring annual‐invaded communities?

Reestablishing native perennial vegetation in annual grass‐invaded rangelands is critical to restoring ecosystems. Control of exotics, often achieved with preemergent herbicides, is essential for successful restoration of invaded rangelands. Unfortunately, desirable species cannot be seeded simultaneously with preemergent herbicide application due to nontarget damage. To avoid this, seeding is commonly delayed at least 1 year. Delaying seeding increases the likelihood that annual grasses will begin reestablishing and compete with seeded species. Activated carbon (AC) can provide preemergent herbicide protection for seeded species because it adsorbs and deactivates herbicides. Previous studies suggest that a cylindrical herbicide protection pod (HPP), containing AC and seeds, allows desired species to be seeded simultaneously with the application of the preemergent herbicide imazapic. Unfortunately, imazapic is only effective at controlling annual grasses for 1–2 years. Indaziflam is a new preemergent herbicide which exhibits longer soil activity, with which HPPs may be useful. To assess this possibility, we evaluated seeding two native species (Wyoming big sagebrush [Artemisia tridentata Nutt ssp. wyomingensis] and bluebunch wheatgrass [Pseudoroegneria spicata (Pursh) Á. Löve]), both incorporated into HPPs and as bare seed, at four application rates of indaziflam in a grow room study. HPPs protected seeded species at low, mid, and high rates of indaziflam. The abundance and size of plants was greater in HPPs compared to bare seed treatments. These results suggest that HPPs can be used to seed native grasses and shrubs simultaneously with indaziflam application.     

Successful cryopreservation of Asian elephant (Elephas maximus) spermatozoa

Reproduction in captive elephants is low and infant mortality is high, collectively leading to possible population extinction. Artificial insemination was developed a decade ago; however, it relies on fresh-chilled semen from just a handful of bulls with inconsistent sperm quality. Artificial insemination with frozen–thawed sperm has never been described, probably, in part, due to low semen quality after cryopreservation. The present study was designed with the aim of finding a reliable semen freezing protocol. Screening tests included freezing semen with varying concentrations of ethylene glycol, propylene glycol, trehalose, dimethyl sulfoxide and glycerol as cryoprotectants and assessing cushioned centrifugation, rapid chilling to suprazero temperatures, freezing extender osmolarity, egg yolk concentration, post-thaw dilution with cryoprotectant-free BC solution and the addition of 10% (v/v) of autologous seminal plasma. The resulting optimal freezing protocol uses cushioned centrifugation, two-step dilution with isothermal 285 m Osm/kg Berliner Cryomedium (BC) with final glycerol concentration of 7% and 16% egg yolk, and freezing in large volume by the directional freezing technique. After thawing, samples are diluted 1:1 with BC solution. Using this protocol, post-thaw evaluations results were: motility upon thawing: 57.2 ± 5.4%, motility following 30 min incubation at 37 °C: 58.5 ± 6.0% and following 3 h incubation: 21.7 ± 7.6%, intact acrosome: 57.1 ± 5.2%, normal morphology: 52.0 ± 5.8% and viability: 67.3 ± 6.1%. With this protocol, good quality semen can be accumulated for future use in artificial inseminations when and where needed.