Reversal of Multidrug Resistance and Inhibition of Phosphorylation of AKT in Human Ovarian Cancer Cell Line by Wild-type PTEN Gene

The reversing effect of wild-type PTEN gene on resistance of C 13K cells to cisplatin and its inhibitory effect on the phosphorylation of protein kinase B （AKT） were studied. The expression of PTEN mRNA and protein in OV2008 cells and C13K cells were semi-quantitatively detected by using RT-PCR and Western blotting. Recombinant eukaryotic expression plasmid containing human wild-type PTEN gene was transfected into C13K cells by lipofectamine2000. The expression of PTEN mRNA was monitored by RT-PCR and the expression of PTEN, Akt, p-Akt protein were ana- lyzed by Western blotting in PTEN-transfected and non-transfected C13K cells. Proliferation and chemosensitivity of cells to DDP were measured by MTT, and cell apoptosis was detected by flow cytometry after treatment with cisplatin. The expression of PTEN mRNA and protein in OV2008 cells were significantly higher than those in C13K cells. After transfection with PTEN gene for 48 h, the expression of PTEN mRNA and protein in C 13K cells were 2.04 ± 0.10, 0.94± 0.04 respectively and the expression of p-Akt protein （ 0.94± 0.07） was lower than those in control groups （1.68 ±0.14, 1.66± 0.10） （P〈 0.05）. The IC50 of DDP to C 13 K cells transfected with PTEN （7.2± 0.3 la mol/L） was obviously lower than those of empty-vector transfected cells and non-transfected cells （12.7±0.4 lamol/1, 13.0±0.3 lamol/L） （P〈0.05）. The apopototis ratio of wild-type PTEN-transfected, empty vector transfected and non-transfected C13K cells were （41.65___0.87）%, （18.61 ±0.70）% and （15.28±0.80）% respectively, and the difference was statistically significant （P〈0.05）. PTEN gene plays an important role in ovarian cancer multidrug resistance. Transfection of PTEN could increase the expression of PTEN and restore drug sensitivity to cisplatin in human ovarian cancer cell line C 13K with multidrug-resistance by decreasing the expression of p-Akt.

Combining QTL mapping and expression profile analysis to identify candidate genes of cold tolerance from Dongxiang common wild rice(Oryza rufipogon Griff.)

Rice（Oryza sativa L.）, a tropical and subtropical crop, is susceptible to low temperature stress during seedling, booting, and flowering stages, which leads to lower grain quality levels and decreasing rice yields. Cold tolerance is affected by multiple genetic factors in rice, and the complex genetic mechanisms associated with chilling stress tolerance remain unclear. Here, we detected seven quantitative trait loci（QTLs） for cold tolerance at booting stage and identified one cold tolerant line, SIL157, in an introgression line population derived from a cross between the indica variety Guichao 2, as the recipient, and Dongxiang common wild rice, as the donor. When compared with Guichao 2, SIL157 showed a stronger cold tolerance during different growth stages. Through an integrated strategy that combined QTL-mapping with expression profile analysis, six candidate genes, which were up-regulated under chilling stress at the seedling and booting developmental stages, were studied. The results may help in understanding cold tolerance mechanisms and in using beneficial alleles from wild rice to improve the cold tolerance of rice cultivars through molecular marker-assisted selection.

Comparative genetic mapping revealed powdery mildew resistance gene MlWE4 derived from wild emmer is located in same genomic region of Pm36 and Ml3D232 on chromosome 5BL

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most devastating wheat diseases. Wild emmer wheat（Triticum turgidum ssp. dicoccoides） is a promising source of disease resistance for wheat. A powdery mildew resistance gene conferring resistance to B. graminis f. sp. tritici isolate E09, originating from wild emmer wheat, has been transferred into the hexaploid wheat line WE4 through crossing and backcrossing. Genetic analyses indicated that the powdery mildew resistance was controlled by a single dominant gene, temporarily designated Ml WE4. By mean of comparative genomics and bulked segregant analysis, a genetic linkage map of Ml WE4 was constructed, and Ml WE4 was mapped on the distal region of chromosome arm 5BL. Comparative genetic linkage maps showed that genes Ml WE4, Pm36 and Ml3D232 were co-segregated with markers XBD37670 and XBD37680, indicating they are likely the same gene or alleles in the same locus. The co-segregated markers provide a starting point for chromosome landing and map-based cloning of Ml WE4, Pm36 and Ml3D232.

Cloning and Sequence Analysis of Disease Resistance Gene Analogues from Three Wild Rice Species in Yunnan

Two sets of degenerate oligonucleotide primers were designed according to amino acid conserved regions of reported plant disease resistance genes which encode proteins that contain nucleotide-binding site and leucine-rich repeats(NBS-LRR), and the plant disease resistance genes which encode serine/threonine protein kinase(STK). By polymerase chain reaction(PCR), disease resistance gene analogues have been amplified from three wild rice species in Yunnan Province, China. The DIN A fragments from amplification have been cloned into the pGEM-T vector respectively. Sequencing of the DNA fragments indicated that 7 classes, 2 classes and 6 classes NBS-LRR disease resistance gene analogues from Oryza rufipogon Griff. , Oryza officinalis Wall. , and Oryza meyeriana Baill. were obtained respectively. The two representative fragments of TO12 from Oryza officinalis Wall, and TR19 from Oryza rufipogon Griff, belong to the same class and homology of their sequences are 100%. The result shows that the sequences of the same class disease resistance gene analogues have no difference among different species of wild rice. 5 classes STK disease resistance gene analogues were also obtained among which 4 classes from Oryza rufipogon Griff. , 1 class from Oryza officinalis Wall. By comparison analysis of amino acid sequences. we found that the obtained disease resistance gene analogues have very low identity(low to 25%) with the reported disease resistance gene L6, N, Bs2, Prf, Pto, Lr10 and Xa21 etc. The finding suggests that the obtained disease resistance gene analogues are analogues of putative disease resistance genes that have not been isolated so far.

马铃薯野生种烯酰水合酶超家族基因ScDHNS的克隆与功能分析

【目的】1,4-二氢氧-2-石脑-CoA合成酶(1,4-dihydroxy-2-naphthoyl-CoA synthase,DHNS)基因是茄科植物糖苷生物碱合成代谢的潜在重要基因,开展马铃薯DHNS基因功能研究与验证,为低糖苷生物碱马铃薯品种(系)的选育提供基因和材料来源。【方法】利用RACE方法克隆得到马铃薯野生种恰柯薯(Solanum chacoense)ScDHNS基因,对其进行生物信息学分析和亚细胞定位,通过构建过表达载体pBWA(V)HS-DHNS转化马铃薯栽培种进行功能验证。【结果】ScDHNS cDNA序列开放阅读框1023 bp,编码340个氨基酸,分子量为37.34 kD,等电点pI为8.592,具有典型的ECH保守结构域,属于烯酰水合酶超家族成员,在二穗短柄草(Brachypodium distachyon)、蒺藜苜蓿(Medicago truncatula)等植物基因组中都有其同源基因,且存在基因扩张和收缩事件。过表达ScDHNS基因后发现转化株ScDHNS和SGT1基因表达量显著上调,且表达量显著高于马铃薯WT植株。且对应转化植株的总糖苷生物碱含量显著高于马铃薯WT植株,最高可达到364.3 mg/kg,是对照的2.4倍。亚细胞定位结果显示ScDHNS定位于过氧化物酶体。【结论】马铃薯ScDHNS基因可能参与调控糖苷生物碱合成关键基因SGT1的表达,通过β-氧化途径和甲羟戊酸通路协同影响糖苷生物碱的合成,该基因与糖苷生物碱在亚细胞水平上的区室化有重要关系,对于培育低糖苷生物碱的马铃薯品种(系)具有重要的应用价值。

基于线粒体COⅠ基因序列的梭鲈野生群体遗传结构

为了解梭鲈种群的遗传结构,实验利用线粒体细胞色素c氧化酶Ⅰ亚基(COⅠ)基因部分序列分析了中国6个和中亚2个群体的遗传差异,并与欧洲群体的单倍型序列进行了比较。结果在640 bp的COⅠ基因序列中检测到5个变异位点,定义了7种单倍型,发现Hap1为8个梭鲈群体的共享单倍型,且与欧洲群体的HapA相同,在中国群体所占比例(93.36%)高于中亚群体(72.58%)和欧洲群体(53.85%);Hap2和Hap3是中国群体的特异单倍型,而Hap4~Hap7为中亚群体的特异单倍型。单倍型序列的聚类图和网络图均显示Hap1/A为梭鲈群体的原始单倍型,中国和中亚群体的特异单倍型相对于原始单倍型仅有1~2个位点的变异,属于Hap1/A的亚型,与欧洲群体的特异单倍型具有较大的差异。每个群体检测到1~4种单倍型,斋桑湖(ZS)群体单倍型最多,而中国的腾格里湖(NX)、兴凯湖(XK)和鸭绿江(YJ)群体仅有1个单倍型(Hap1);塔什干(TS)群体的单倍型多样性(Hd)和核苷酸多样性(π)最高(Hd=0.514±0.069;π=0.00079±0.00011),其次是ZS群体,而中国梭鲈群体的多样性参数较低。AMOVA分析结果显示,梭鲈群体间遗传变异占20.74%,群体间遗传分化程度较高(0.15≤F_(st)=0.20736<0.25),TS群体与ZS群体和中国群体间的遗传分化极大(F_(st)>0.25),中国群体中仅黑河(HH)群体与其他群体的遗传分化较大,而中国其他5个群体间无遗传分化。基于群体间遗传距离的系统进化树显示,来自中国的6个梭鲈群体与哈萨克斯坦的ZS群体聚为一支,而乌兹别克斯坦的TS群体独立为一支。研究结果为梭鲈群体的繁殖及放流管理提供了参考。

Identification of quantitative trait loci and candidate genes associated with ABA sensitivity in common wild rice (Oryza rufipogon Griff,)

Abscisic acid （ABA）, as one of the foremost signaling molecules in plants, is an important hormone which plays versatile functions in regulating developmental process and adaptive stress process. A set of introgression lines were previously generated via a backcrossing program using an elite indica cultivar rice Teqing （O. sativa L.） as recipient and an accession of Yuanjiang common wild rice （O. rufipogon Griff.） as donor. In this study, the previously developed introgression lines were evaluated for ABA sensitivity. Here we reported that a total of 14 quantitative trait loci （QTLs） associated with ABA sensitivity were identified. An ABA sensitive introgression line, YIL53, was identified and characterized. Physiological characterization, including chlorophyll content, malondialdehyde content, soluble sugar content, and stomata movement, demonstrated that YIL53 exhibited the characteristics associated with ABA sensitivity. Genotypic analysis revealed that YIL53 harbored one QTL related to ABA sensitivity, qASS1-2, which was located on chromosome 1 within one introgressed segment derived from the Yuanjiang common wild rice. Furthermore, the qASS1-2 was finally narrowed down to a 441-kb region between simple sequence repeats （SSR） marker RM212 and single nucleotide polymorphism （SNP） marker M3 using the segregation population derived from the cross between Teqing and YIL53, and three candidate genes associated with ABA sensitivity were identified using a strategy combined gene expression analysis with QTL mapping. Identification of the QTLs related to ABA sensitivity and characterization of the ABA sensitive line YIL53 would provide a helpful basis for isolating novel genes related to ABA sensitivity.

喷施水杨酸对野生大豆草甘膦耐性的影响

为进一步提高前期筛选到的高耐草甘膦野生大豆材料的抗性,本研究以喷施清水为对照,先后对野生大豆材料进行外源喷施水杨酸(SA)和草甘膦处理,测定过氧化氢(H_(2)O_(2))、超氧阴离子(O_(2)^(-))、莽草酸含量、脂质过氧化水平(丙二醛含量)、叶绿素含量、抗氧化酶活性及5-稀醇式丙酮-莽草酸-3-磷酸合成酶(EPSPS)基因表达等指标。结果表明,与仅喷施草甘膦阳性对照相比,喷施0.5 mmol·L^(-1) SA的2个野生大豆材料H_(2)O_(2)、O_(2)^(-)、莽草酸和丙二醛含量在草甘膦处理后整体显著降低,叶绿素含量、过氧化物酶(POD)、过氧化氢酶(CAT)和超氧化物歧化酶(SOD)活性整体显著升高,EPSPS基因表达整体显著上调。上述结果表明,外源喷施SA可提高野生大豆植株过氧化物清除酶活性,进一步提高植株对草甘膦的耐性水平。本研究为提高栽培大豆草甘膦耐性,降低草甘膦对大豆植株损伤提供了参考。

野生大豆功能基因的研究进展

野生大豆(Glycine soja Sieb.&Zucc.)是栽培大豆(Glycine max [L.] Merr.)的近缘祖先种。在大豆驯化的过程中,栽培大豆丢失了大量的基因或等位变异,导致栽培大豆的遗传多样性降低,这严重限制了栽培大豆品种选育和改良的有效性与丰富性。我国野生大豆种质资源丰富,蕴藏着许多高蛋白含量、抗病虫、耐干旱、耐盐碱等方面的潜力基因,挖掘潜力基因并利用分子设计育种技术应用到现代的栽培大豆品种中,能够有效地拓宽栽培大豆的遗传多样性。本文综述了野生大豆的分布规律和形态特征、近年来在野生大豆中发掘的重要功能基因或位点,包括百粒重、开花期和成熟期、蛋白质和油分含量、抗病、抗虫、耐盐碱、耐干旱等重要农艺性状基因,并讨论这些重要基因或位点在未来栽培大豆育种中的应用潜力,以期为育种家培育和改良大豆新品种提供一种新的育种思路和策略。

野生大豆与栽培大豆几丁质酶基因鉴定及其表达分析

几丁质酶是以几丁质等为底物的糖基水解酶(GH,glycosyl hydrolases),在植物生长发育及抵御逆境中发挥重要功能。然而,大豆几丁质酶基因的组织表达模式及对逆境的响应尚不清楚,影响了其在遗传改良中的应用。本研究分别鉴定野生大豆与栽培大豆几丁质酶基因,并分析其表达模式。结果发现,野生大豆与栽培大豆分别含62个和55个几丁质酶基因,位于17条和18条染色体上;进化树分析发现,117个基因分为5类,其中类群Ⅲ与类群Ⅴ属于GH18亚家族,类群Ⅰ、类群Ⅱ与类群Ⅳ属于GH19亚家族;启动子分析发现,几丁质酶家族成员包含响应激素及逆境胁迫顺式作用元件。栽培大豆几丁质酶基因表达分析发现,其在不同组织及抗病耐逆等过程差异表达,其中Gm.01G142400和Gm.13G346700等在接种花叶病毒的抗病品种叶片中诱导表达,Gm.03G254300和Gm.20G164600等在低磷胁迫的磷高效品种根系中诱导表达,Gm.08G259200和Gm.19G245400等在低磷胁迫根瘤中诱导表达;野生大豆几丁质酶基因表达分析发现,其在不同组织及耐盐过程差异表达,其中Gs.02G002604和Gs.02G002940等在盐胁迫耐盐品种叶片中诱导表达。以上结果为挖掘利用几丁质酶基因奠定了基础。

近40年云南普通野生稻优异特性及有利基因的发掘与利用

稻种资源是栽培稻育种的物质基础,也是研究栽培稻起源、进化、发育和基因功能的重要对象。在众多野生稻中,普通野生稻与栽培稻亲缘关系最近,是培育栽培稻新品种的优异基因供体,因此了解清楚其遗传背景尤为重要。以云南普通野生稻为对象,对其遗传背景的研究和有利基因的运用进行总结和分析,首先详细梳理了近年来从云南普通野生稻资源中发掘出的优异特性,阐述了其各种优良性状;其次系统总结了云南普通野生稻有利基因的发掘现状,囊括了目前从中发掘出来的重要功能基因;然后综述了云南普通野生稻优异特性在创制新种质方面所取得的成就,包括滇9型籼型不育系的育成及大量育种中间材料的获得;最后针对云南普通野生稻开发研究的优势和存在的问题,提出今后的研究重点以及有利基因发掘利用的建议,并对其利用的潜力进行了展望,以期为云南普通野生稻的进一步研究、开发和利用提供理论支持。

野生鸟类介导抗生素抗性基因传播的研究进展

细菌的抗生素耐药是重大的公共卫生问题,抗生素抗性基因(antibiotic resistance genes, ARGs)可能会在人类、动物和环境微生物组之间水平转移,使其相互依存的生态系统与公共卫生之间存在潜在关联性。鸟类是地球上种类繁多的物种之一,它们的跨距离传播可能会加速耐药基因在全球范围内的扩展,带来一定的健康风险。本文综述了野生鸟类介导抗生素抗性基因在生态环境中的主动传播,提出其作为传播媒介的重要性,并分别阐述了城市留鸟和候鸟对抗生素抗性基因的传播作用,着重突出了候鸟跨地区传播的作用,总结出野生鸟类介导抗生素抗性基因的传播是尤为重要的研究方向。

长沙生态动物园野生动物衣原体感染情况调查

为了解长沙生态动物园野生动物衣原体的感染情况,本试验根据中华人民共和国农业行业标准(NY/T 562—2015),采用巢式PCR方法对采集到的园区内63种动物的231份粪便样品进行衣原体检测,并对阳性样品进行测序分析。结果显示,所有检测样品中有13份样品呈衣原体阳性,总阳性率为5.63%(13/231),其中植食类动物阳性率为11.43%(4/35)、灵长类动物阳性率为13.95%(6/43)、禽鸟类动物阳性率为3.13%(2/64)、肉食类动物阳性率为1.12%(1/89)。所有动物中感染风险最大的是灵长类动物,11种灵长类动物中有5种检出阳性,占比45.45%,且优势比(OR)=4.19(P<0.01)。基因测序和比对结果显示,仅从1份绿孔雀阳性粪便样品中获得有效的衣原体序列(序列号:OK033359.1),其与美国分离株c149(序列号:KY206900.1)相似度达99.72%,可鉴定为Chlamydia gallinacea,且处于较早进化阶段。结果表明,该动物园存在衣原体感染,但感染率不高,感染风险最大的是灵长类动物,禽鸟类存在Chlamydia gallinacea感染。衣原体在生态动物园野生动物中的存在对人类健康存在潜在威胁,应引起重视。

Conventional and Molecular Approaches towards Genetic Improvement in Pigeonpea for Insects Resistance

Pigeonpea [Cajanus cajan (L.) Millspaugh] is an important food legume of the semi-arid tropics (SAT) sustaining livelihood of millions of people. Stagnant and unstable yield per hectare all over the world is the characteristic feature of this crop. This is primarily ascribed to its susceptibility/sensitivity to a number of biotic and abiotic factors. Among biotic factors, insects such as pod borer (Helicoverpa armigera), pod fly (Melanoagromyza obtusa) and spotted borer (Maruca vitrata) substantially damage the crop and result in significant economic losses. Management of these insects by genetic means has always been considered environment friendly approach. However, genetic improvement has always been impeded by limited genetic variability in the primary gene pool of pigeonpea. Wild species present in the secondary and tertiary gene pools have been reported to carry resistance for such insects. However, transfer of resistance through conventional backcrossing has not been much successful. It calls for gene introgression through marker assisted backcrossing (MABC) or advanced backcross breeding (AB breeding). In this review, we have attempted to assess the progress made through conventional and molecular breeding and suggested the ways to move further towards genetic enhancement for insects resistance in

Brown Planthopper Resistance Genes in Rice:from Germplasm to Breeding

The brown planthopper (BPH), Nilaparvata lugens Stal (Homoptera: Delphacidae), is one of the most destructive and widespread insect pests of rice (Oryza sativa) that can be found throughout the rice-growing areas

Differential RNA Editing of Mitochondrial Genes in WA-Cytoplasmic Based Male Sterile Line Pusa 6A, and Its Maintainer and Restorer Lines

RNA editing changes the nucleotides at the transcript level of mitochondrial genes which results in synthesis of functional proteins.This study was designed to find the editing sites which could be implicated in male fertility restoration and to develop editing based markers for differentiation of cytoplasmic male sterility and maintainer lines from each other.DNA and RNA from young panicles were isolated from three-line system of hybrid rice PRH10,wild abortive(WA)cytoplasm based male sterile(A line Pusa 6A),maintainer(B line Pusa 6B)and restorer(R line PRR78)lines.Pusa 6A and PRR78 having the same WA cytoplasm are allo-nuclear and iso-cytpolasmic lines.The genomic and cDNA amplicons for eight mitochondrial genes(18SrRNA,atp6,atp9,cobII,coxI,coxIII,nadI and rps3)were sequenced and compared.Differences in genomic and cDNA sequences were considered as editing.Two hundred and thirty editing sites having base substitution or insertion/deletion were identified with the highest in 18SrRNA(5.74%)and the lowest in coxI(0.60%).The highest editing sites were observed in fertile maintainer Pusa 6B followed by PRR78 and Pusa 6A,of which random five editing sites in five different rice mitochondrial transcripts namely atp9,cobII,coxIII,rps3 and 18SrRNA were chosen and validated through cleaved amplified polymorphism sequence(CAPS)analysis and found to be partially edited in four genes.The identical editing sites of different mitochondrial genes from maintainer and restorer lines might reflect their possible contribution to fertility restoration of sterile WA cytoplasm.

Comparative studies on some genetic characteristics among four large yellow croaker (Pseudosciaena crocea) populations

The genetic structure was compared among four large yellow croaker （Pseudosciaena crocea） populations, an important aquaculture species in China. RAPD and ISSR results show that the genetic distances between domestic stocks and their wild counterparts are significantly larger than those between the three wild stocks. The sequence of mtDNA cyt b shows that the three wild populations share the same sequence, with differences in the four bases from the cultured stock： two transitions [57（ C-T）, 291 （G-A） ] and two transversions [66（C-G） ,223（A-C） ]. Cluster analysis reveals that the domestic stock has diverged from its parental wild stock [ Min-Yuedong （ Fujian Province-eastern Guangdong Province, China） stock]. Results demonstrate that 20 a of domestication of the Pseudosciaena crocea stock has resulted in significant genetic changes.

能谱CT成像预测肺腺癌EGFR突变的价值研究

目的 分析能谱CT成像与肺腺癌表皮生长因子受体(epidermal growth factor receptor,EGFR)突变状态之间的关系。方法 纳入2019年1月—2022年12月在福建医科大学附属协和医院收治的肺腺癌且对EGFR突变进行检测者120例,根据EGFR检测状态分为EGFR野生型(n=66)与EGFR突变型(n=54)。定量测量能谱CT的能谱曲线计算曲线斜率(curve slope,λHU)、水浓度、碘浓度、有效原子序数、CT 70 keV、CT 40 keV等定量参数指标,分析两组的能谱CT定量参数指标变化。结果 EGFR野生型的吸烟史、性别与EGFR突变型相比,差异有统计学意义(P <0.05);EGFR突变型在能谱CT静脉期、动脉期的定量参数包括λHU、有效原子序数、碘浓度、CT 70 keV值、CT 40 keV值等定量参数与EGFR野生型相比大幅度增高,差异有统计学意义(P <0.05);两组水浓度相比,差异无统计学意义(P> 0.05)。结论 能谱CT成像定量测量对肺腺癌表皮生长因子受体突变状态的预测具有较高的应用价值,可用于表皮生长因子受体的突变状态的预测诊断。

基于P72、MGF和CD2v基因的非洲猪瘟三重荧光定量PCR检测方法的建立

非洲猪瘟在临床上出现多种毒株,主要分为野毒株和基因变异毒株。因其在临床上造成不同程度的危害,有必要建立一种方法进行正确诊断和健康监测。为实现野毒株和基因变异毒株的鉴别诊断,研究根据GenBank中的ASFV中国流行株(登录号:MK333180.1)的P72、CD2v、MGF基因保守序列设计引物,建立一种可以同时检测上述三种基因的三重荧光定量PCR方法。三重荧光定量PCR方法对猪细小病毒(PPV)、猪伪狂犬病病毒2型(PRV)和猪圆环病毒(PCV2)检测均无交叉反应,表现出较强的特异性。敏感性检测显示针对P72、CD2v和MGF重组质粒标准品的最低检测下限为103~102 copies/mL。在重复性试验中,大部分组间变异系数均小于2%或接近2%,具有较好的重复性。试验表明,三重荧光定量PCR方法可用来鉴别野毒株和基因变异毒株。

新疆野生核桃JfDREB1A基因的克隆与原核表达分析

为探究DREB1A基因在新疆野生核桃响应低温胁迫中的作用及表达特性,以新疆野生核桃群体中的‘小矩圆’类型为试验材料,根据同源克隆的方法得到JfDREB1A基因的cDNA全长序列,利用荧光定量PCR检测其在低温胁迫条件下的表达水平;将该基因片段重组到pET-28a(+)原核表达载体上,转化到大肠杆菌(DE3)感受态细胞中诱导表达。结果表明,获得了新疆野生核桃中的DREB1A,将其命名为JfDREB1A,该基因的开放读码框长度为645 bp,具有典型的DREB1/CBF转录因子结构特征,JfDREB1A蛋白与其他植物DREB1蛋白具有高度保守性,与核桃亲缘关系最近。RT-PCR分析显示在4℃低温胁迫下,JfDREB1A基因的表达量迅速上调,8 h时达到最大。成功构建了pET-28a-JfDREB1A重组表达质粒,得到大小约为29 ku的融合蛋白,与预测蛋白大小相符,该蛋白在大肠杆菌中最佳诱导条件是诱导剂IPTG浓度为0.5 mmol/L,诱导2 h。