云南省角蝉科二新属二新种

最近我们从全国动物学科研单位和高等院校借调来大批的角蝉科标本,进行系统的分类研究,从中发现有二新属及二新种,均采自云南西双版纳,现在先把它们加以记载。     

Reagents for Modification of Protein–Nucleic Acid Complexes: III.1Site-Specific Photomodification of Elongation Complex of DNA Polymerase β with Arylazide Derivatives of Primers Sensitized with Fluorescent ATP γ-Amides

ATP -amides containing in -N-position 1-methylpyrene, 9-methylanthracene, 10-chloro-9-methyl-anthracene, and 3-methylperylene residues were synthesized and characterized. These compounds were used as sensitizers of site-specific photomodification of the reconstituted elongating complex of the mammalian DNA polymerase . The photomodification was carried out with the use of photoaffinity reagents, which were synthesized in situby the 5"-³²P-labeled primers extension with photoreactive analogues of dCTP containing in the exo-N-position of cytosine various perfluoroarylazide groups. The effect of structures of the sensitizers and photoreactive reagents on the efficiency and selectivity of photocrosslinking of primers to the enzyme and template, as well as formation of a number of other photomodification products was studied. It was shown that the sensitizers containing 10-chloro-9-methylanthracene and 3-methylperylene residues allow one to obtain photocrosslinks under such irradiation conditions when photomodification in their absence is not essentially observed.     

RPA repair recognition of DNA containing pyrimidines bearing bulky adducts

Recognition of new DNA nucleotide excision repair (NER) substrate analogs, 48-mer ddsDNA (damaged double-stranded DNA), by human replication protein A (hRPA) has been analyzed using fluorescence spectroscopy and photoaffinity modification. The aim of the present work was to find quantitative characteristics of RPA-ddsDNA interaction and RPA subunits role in this process. The designed DNA structures bear bulky substituted pyrimidine nitrogen bases at the inner positions of duplex forming DNA chains. The photoreactive 4-azido-2,5-difluoro-3- pyridin-6-yl (FAP) and fluorescent antracenyl, pyrenyl (Antr, Pyr) groups were introduced via different linker fragments into exo-4N of deoxycytidine or 5C of deoxyuridine. J-dU-containing DNA was used as a photoactive model of undamaged DNA strands. The reporter group was a fluorescein residue, introduced into the 5'-phosphate end of one duplex-forming DNA strand. RPA-dsDNA association constants and the molar RPA/dsDNA ratio have been calculated based on fluorescence anisotropy measurements under conditions of a 1:1 RPA/dsDNA molar ratio in complexes. The evident preference for RPA binding to ddsDNA over undamaged dsDNA distinctly depends on the adduct type and varies in the following way: undamaged dsDNA < Antr-dC-ddsDNA < mmdsDNA < FAPdU-, Pyr-dU-ddsDNA < FAP-dC-ddsDNA (K(D) = 68 +/- 1; 25 +/- 6; 13 +/- 1; 8 +/- 2, and 3.5 +/- 0.5 nM correspondingly) but weakly depends on the chain integrity. Interestingly the bulkier lesions not in all cases have a greater effect on RPA affinity to ddsDNA. The experiments on photoaffinity modification demonstrated only p70 of compactly arranged RPA directly interacting with dsDNA. The formation of RPA-ddsDNA covalent adducts was drastically reduced when both strands of DNA duplex contained virtually opposite located FAP-dC and Antr-dC. Thus RPA requires undamaged DNA strand presence for the effective interaction with dsDNA bearing bulky damages and demonstrates the early NER factors characteristic features underlying strand discrimination capacity and poor activity of the NER system toward double damaged DNA.     

角蝉科一新属一新种

角蝉科的昆虫,是世界著名的富于拟态性的昆虫。它们的畸形发达的前胸背板给人们以明显的识别的特征。这一科不少种类是果树、森林或农业的重要害虫,造成相当严重的灾害。角蝉科截至目前为止,全世界已记载的共324属2500余种;中国已知的20属152种。 本文记述角蝉科一新属及一新种。     

中国的广翅蜡蝉科附八新种

广翅蜡蝉科的种类,多数为害木本植物,少数为害禾本科草本植物,有的是果树、森林和农作物的害虫。但关于广翅蜡蝉科的研究,过去很不充分,我国的种类、分布与寄主关系,至今还不清楚。1935年胡经甫的昆虫名录中列入11种(包括3种同物异名),1959年《中国果树病虫志》中列了5种。 我们根据三十年来在全国各地的采集调查,和全国兄弟单位提供的标本,共鉴定出26种。本文着重记录它们的分布及其为害植物,并记载新种8种。     

金翅夜蛾亚科八新种及一些种类的订正

金翅夜蛾亚科的种类,有的在外形上非常相似,因之过去文献记载常有混淆的地方和鉴定上的错误。本文讨论了一些种类的区别,纠正某些错误的记载,并报道8个新种。 所有新种的模式标本都暂在西北农学院昆虫标本室保存。     

Highly selective affinity labeling of DNA-polymerase from Thermus thermophilus B35 by a binary system of photoreactive agents

The thermostable DNA-polymerase from Thermus thermophilus B35 (Tte-polymerase) was affinity labeled by a binary system of photoreagents comprising base-substituted TTP analogs. The 5;-[32P]-labeled primer was elongated by Tte-polymerase in the presence of a TTP analog containing the photoreactive 2,3,5, 6-tetrafluoro-4-azidobenzoyl group (FAB-4-dUTP). Then the reaction mixture was UV-irradiated (365-450 nm) in the presence or the absence of a photosensitizer (TTP analog containing a pyrene moiety, Pyr-dUTP). The initial rate of the Pyr-dUTP-sensitized photomodification was almost 10-fold higher than the rate of direct photomodification (in the absence of Pyr-dUTP); in the case of the sensitized modification, the product of covalent cross-linking of the photoreactive primer with Tte-polymerase was apparently homogenous according to the data of electrophoresis. The enzyme was protected from the photosensitized modification by dNTP. To confirm the selectivity of the photosensitized modification of Tte-polymerase, another DNA-binding protein (human replication factor A, RPA) was added to the reaction mixture. In the presence of the photosensitizer (Pyr-dUTP), RPA was not labeled and only Tte-polymerase was modified, whereas in the case of direct modification, Tte-polymerase and the p32 and p70 subunits of RPA were labeled. The suggested method enables highly selective affinity modification of DNA-polymerases.     

Interaction of Human Replication Protein A with DNA Duplexes Containing Gaps of Varying Size

Replication protein A (RPA) is a heterotrimeric protein that has high affinity for single-stranded (ss) DNA and is involved in DNA replication, repair, and recombination in eukaryotic cells. Photoaffinity modification was employed in studying the interaction of human RPA with DNA duplexes containing various gaps, which are similar to structures arising during DNA replication and repair. A photoreactive dUMP derivative was added to the 3" end of a gap-flanking oligonucleotide with DNA polymerase , and an oligonucleotide containing a 5"-photoreactive group was chemically synthesized. The 5" end predominantly interacted with the large RPA subunit (p70) regardless of the gap size, whereas interactions of the 3" end with the RPA subunits depended both on the gap size and on the RPA concentration. Subunit p32 was mostly labeled in the case of a larger gap and a lower RPA concentration. The results confirmed the model of polar RPA–DNA interaction, which has been advanced earlier.     

金翅夜蛾亚科的研究

近年来,我们又获得大量金翅夜蛾亚科的标本。这些标本有的是我们区系组的同志从全国各地采集的,有的是兄弟单位赠送或借给的。 标本共77,085号,我们逐一进行了外部特征的观察,每种作了雌雄生殖器的解剖研究。除前一报告已记载的相同种类外,新增加的种类有36种。如下: