A revolutionary tool: CRISPR technology plays an important role in construction of intelligentized gene circuits

With the development of synthetic biology, synthetic gene circuits have shown great applied potential in medicine, biology, and as commodity chemicals. An ultimate challenge in the construction of gene circuits is the lack of effective, programmable, secure and sequence‐specific gene editing tools. The clustered regularly interspaced short palindromic repeat (CRISPR) system, a CRISPR‐associated RNA‐guided endonuclease Cas9 (CRISPR‐associated protein 9)‐targeted genome editing tool, has recently been applied in engineering gene circuits for its unique properties‐operability, high efficiency and programmability. The traditional single‐targeted therapy cannot effectively distinguish tumour cells from normal cells, and gene therapy for single targets has poor anti‐tumour effects, which severely limits the application of gene therapy. Currently, the design of gene circuits using tumour‐specific targets based on CRISPR/Cas systems provides a new way for precision cancer therapy. Hence, the application of intelligentized gene circuits based on CRISPR technology effectively guarantees the safety, efficiency and specificity of cancer therapy. Here, we assessed the use of synthetic gene circuits and if the CRISPR system could be used, especially artificial switch‐inducible Cas9, to more effectively target and treat tumour cells. Moreover, we also discussed recent advances, prospectives and underlying challenges in CRISPR‐based gene circuit development.     

塔克拉玛干沙漠腹地人工植被下土壤微生物的初步研究

在极端干旱的塔克拉玛干沙漠腹地利用矿化度为4～5 g/L的地下咸水灌溉,在风沙土上建立了人工绿地。对流沙和人工绿地中微生物三大类群（细菌、真菌和放线菌）进行了数量测定,结果显示：流沙上微生物数量很少,人工绿地的微生物数量显著增加;种植时间相同而植被类型不同的样地中的微生物数量有差异;微生物的组成中,细菌占绝大多数,达微生物总数的90%以上,其次是放线菌,真菌数量最少;表层微生物数量明显高于下层。人工绿地微生物数量的变化,说明风沙土已逐步向具有一定肥力水平的土壤方向演变。     

家马的驯化起源与遗传演化特征

人类文明发展历史中, 家马(Equus ferus caballus)曾是推动文化交流、促进人类社会发展的主要动力。关于家马何时、何地被驯化以及在此过程中其遗传演化如何被人类影响等一直备受关注。近年来随着遗传学技术的发展, 人们对该问题有了更为深入的理解。本文回顾了近二十年来相关研究所取得的成果, 探讨了家马的驯化起源中心和驯化过程中的遗传演化特征, 并对未来的研究方向以及遗传资源保护提出了建议。分子标记遗传学和考古学研究认为家马可能来自多个驯化起源地种群, 然而最近的古DNA研究结果表明, 现代家马的驯化起源可能比之前人们所猜测的更加复杂, 古代博泰马被认为是最早被驯化的家马, 然而最近被证实并不是现代家马的直系祖先。如此复杂的驯化问题可能从多学科的层次才能解析清楚。人类社会活动直接或间接影响了家马的演化历程, 特别是工业革命以来家马的遗传基础发生了巨大变化, 其遗传多样性开始急剧衰退, 不少地方品种正逐渐走向衰落甚至灭绝。为确保农业生态安全不受威胁, 建议加强家马遗传资源保护与动物遗传学和文化地理之间的联系研究。     

腾格里沙漠沙坡头地区荒漠地衣与生物地毯工程

文章论述了荒漠地衣与“沙漠生物地毯工程”。在沙坡头结皮微型生物中发现了23种地衣,其中两个新种已发表,一属6种为中国新记录。对于在腾格里沙漠东南角沙坡头地区人工植被固沙防护体系建成后的生态演替进行了分析。由于人工植被为形成结皮的微型生物提供了适宜的生长环境而导致微型生物结皮的形成和发育。在水分平衡规律的作用下漫长生态演替过程中,具有抽水机效应的人工植被使沙土深层水分消耗殆尽,从而导致人工植被自身逐年衰退。然而,与此相反的是无抽水机效应而具有固沙、固碳和抗旱功能的结皮微型生物却逐年形成并发育。这一结果为借助于结皮微型生物的接种技术在干旱沙漠构建“沙漠生物地毯工程”的可行性提供了科学依据。为了优化“沙漠生物地毯工程”利用荒漠地衣耐旱基因以构建转基因草地植物的研究也正在进行中。该研究是“沙漠生物地毯工程”基础研究的组成部分。     

山羊瘤胃微生物宏基因组BAC文库的构建与分析

从山羊瘤胃液中提取混合微生物DNA,经BamHI部分酶切得到50kb～800kb的DNA片段后,将其连接到pCCIBAC载体上,转化E．coliEPI300,建立山羊瘤胃微生物BAC文库。经RFLP鉴定分析,该文库12672个克隆,平均插入片段为6lkb。该文库的构建为后续新型基因的筛选提供了材料,为进一步研究山羊瘤胃微生物奠定了基础。     

菱斑巧瓢虫替代饲料的研究

瓢虫成虫饥饿24h后,通过室内对菱斑巧瓢虫Oenopia conglobata (Linnaeus)成虫饲喂24种人工饲料,研究菱斑巧瓢虫的取食行为;以桃蚜(MP)作对照,选择羊肝+蔗糖(SLS)、猪肝+蔗糖(PLS)、黄粉虫蛹+蔗糖(TMPS)3种饲料研究瓢虫产卵量;以白蛾周氏啮小蜂蛹(CCP)、SLS、PLS和MP对瓢虫成虫进行交叉饲喂试验,每周期7d,研究饥饿后恢复产卵时间和产卵量的差异;最后以CCP和MP饲喂瓢虫幼虫,观察其发育历期。结果表明:在羊肝、鸡心、菜蛾幼虫、鸡肝和黄粉虫蛹组中,无论是否添加蔗糖,瓢虫取食情况较好。瓢虫成虫饲喂SLS、TMPS、PLS及MP后,MP组的14d单雌产卵量为166.25粒、SLS组为18.15粒、PLS为9.65粒和TMPS组3.90粒,瓢虫恢复产卵的时间也有差异,MP组仅需要1d,而其它组需要3d至至5d。交叉饲喂试验发现,CCP组产卵量(40.67～46.87粒/雌)明显高于常用饲料-PLS组(4.53～7.13粒/雌),但与MP组(61.80～82.07粒/雌)仍有一定差距。利用CCP饲喂瓢虫幼虫,可以完成整个发育历期,但是存活率较低,1～4龄幼虫各龄期内存活率在63.64%～87.50%之间,并且发育历期(21.23d)明显长于饲喂MP组(15.71d)。     

城市小型浅水人工湖泊浮游藻类与水质特征研究

研究了广州城区某富营养化小型浅水人工湖泊浮游藻类和水质特征及其变化规律。根据镜检共鉴定出7门54属浮游藻类,以绿藻门、蓝藻门、硅藻门和裸藻门为主,其中绿藻门的绿球藻目占绝对优势,达26属62种,占总属数的45.6%;在整个观测期内,浮游藻类密度均较高,05年2月藻类密度高达10.8×106cell·mL-1;根据Kolkwitz划分的水域类型,该湖泊中浮游藻类属于α、β-中污带指示种类的最多。由以修正的卡森(Carlson)指数为基础的综合营养型评价方法和Margalef生物多样性指数判断,该湖泊水体属重富营养化水平,受到重污染,且污染水平季节变化明显:冬季>秋季>夏季。     

An Artificial Protein with the Biological Activity of the Differentiation Factor for the HL-60 Cell Line of Human Promyelocyte Leukemia

The ABB-df artificial protein was prepared by inserting the TGENHR biologically active peptide corresponding to the 41–46 sequence of the differentiation factor for the HL-60 cell line of the human promyelocyte leukemia into the N-terminus of the polypeptide chain of albebetin, an artificial protein with the preset structure. The ABB-df protein was found to induce the differentiation of HL-60 cells and to inhibit their proliferation; its efficiency was almost the same as that of the starting peptide. According to CD spectroscopy, the inclusion of the peptide fragment into albebetin exerts virtually no effect on the regular secondary structure of albebetin.     

复乳法制备红细胞代用品

以单甲氧基聚乙二醇聚乳酸共聚物 [methoxypoly(ethyleneglycol) poly DL lactide,PELA]为膜材 ,用W /O/W的复乳 溶剂扩散法制备了包埋牛血红蛋白 (bovinehemoglobin ,BHb)的微胶囊作为人造红细胞。研究发现复乳过程搅拌分散速率、有机溶剂种类和固化方法对BHb活性有明显影响。当搅拌分散速率小于 90 0 0r/min、以乙酸乙酯为有机相时 ,采用复乳 溶剂扩散法包埋BHb的过程对BHb活性无明显影响。当搅拌分散速率高于 12 0 0 0r/min时 ,BHb活性降低。复乳 溶剂扩散法制备微胶囊过程中固化液体积与微胶囊中BHb活性密切相关 ,通过增大固化液和复乳液的体积比可较好地保持BHb的活性。最后制得了粒径 10 μm左右、包埋率 93%、P50 和Hill系数均接近于天然BHb的微胶囊。     

Genome-wide screening of severe male factor infertile patients using BAC-array comparative genomic hybridization (CGH)

Male factor infertility is present in up to 50% of infertile couples, making it increasingly important in their treatment. Although most research into the genetics of male infertility has focused on the Y chromosome, male factor infertility may result from other genetic factors. We utilized the whole genome array comparative genomic hybridization (CGH) to identify novel genetic candidate associated with severely impaired spermatogenesis. We enrolled 37 patients with severe male factor infertility, defined as severe nonobstructive type oligozoospermia (≤5×10(6)/ml) or azoospermia, and 10 controls. Routine cytogenetic analyses, Yq microdeletion PCR test and whole genome bacterial artificial chromosome (BAC)-array CGH were performed. Array CGH results showed no specific gains or losses related to impaired spermatogenesis other than Yq microdeletions, and there were no novel candidate genetic abnormalities in the patients with severe male infertility. However, Yq microdeletions were detected in 10 patients. Three showed a deletion in the AZFb-c region and the other 7 had deletions in the AZFc region. Although we could not identify novel genetic regions specifically associated with male infertility, whole genome array CGH analysis with higher resolution including larger numbers of patients may be able to give an opportunity for identifying new genetic markers for male infertility.