. from the direct mutant of spirulina platensis ( sp - d ) , we got high purity and activity phycobiliprotein which could grow crystals . the algae fluorescent probe prepared by coupling the above polyclonal antibody to phycobilipotein not only keeps the property of stronger anti - fluorescence quenching but also has the lower fluorescent background when it was used for labeling stoma cells of pea tendril 以原核表达的peac1为抗原制备了免疫活性较好的抗豌豆肌动蛋白的多克隆抗体，从螺旋藻中纯化了高纯度、高活性、能结晶的藻胆蛋白，将两者偶联制备的藻荧光探针，不仅保持了藻胆蛋白很强的抗荧光淬灭能力，而且用于豌豆卷须气孔细胞荧光标记时有更低的荧光背景。
A 1 . 5 kb apramycin resistance fragment was inserted into nru i site of aved gene and the inactivated aved gene fragment was then introduced into mcs region of phjl401 - an e . coli / streptomyces shuttle vector with conjugation function ( containing orit gene ) . as a result of above procedures , a recombinant plasmid pid03 was obtained 将1 . 5kb的安普霉素抗性基因片段插入到aved基因中的nrui酶切位点，再将此灭活的aved基因片段插入到具有接合转移功能(含有orit基因)的链霉菌?大肠杆菌穿梭质粒phjl401的多克隆位点区，由此得到重组质粒pid03 。
The interest gene was inserted in the - tha l . tho 1 multiple cloning sites of donor plasimd pfastbachtb of baculovirus expression system . after analysis by restriction endonuclease and pcr , the recombinant donor plasmid gpl - fast was transforn1ed to the competent celi dhi0bac which contalns the bacmid and the helper plasmid , the recombinant bacmid gpl - bac was acquired which would express the vpl of gpv strain h l 同时将该目的基因插入到杆状病毒表达系统的供体质粒pf _ ( ast ) b _ ( ac ) htb的xba 、 xho多克隆位点间，经酶切、 pcr鉴定后，将重组的供体质粒gp1 - fast转化到含有杆状病毒和辅助质粒的dh10b _ ( ac )感受态细胞中，获得了表达gpvh1株vp1的重组杆状病毒gp1 - bac 。
The gd and ge gene was subcloned into puc18 , resulting in pugdge . the fragment from pcdnas . 1 - including hcmv promoter / enhancer , mcs and neomycin resistance gene was inserted into the bamhi and bsteii restrication sites of pugdge , resulting in the universal transfer vector pgd - m - ge . the universal transfer vector pgd - m - ge has deleted the gi gene and 363bp in the 5 ' end of the ge orf of prv . there were 11 restrication sites for insertion of the foreign gene . the upstream and downstream flanking sequences were up to 1 . 25kb and 1 . 42kb . it will be useful for developing the recombinant prv expressing foreign gene ( s ) 将gd 、 ge基因连接于质粒puc18获得pugdge ，缺失质粒pugdge的bamh和bste位点间391bp的片段。在此缺失位置插入来自质粒pcdna3 . 1 -的一伪狂犬病病毒gd 、 ge 、 tk基因的克隆与通用转移载体的构建段含hcmv启动子。多克隆位点和neo报告基因的片段，构建了通用转移载体ppd m pe 。
Based on a 3 . 1kb pst i fragment of genomic dna of a wild s . avermitilis , a 1 . 5 kb apramycin resistance fragment was inserted into sph i site of avec gene in the 3 . 1 kb fragment , then a recombinant plasmid pc05 was obtained by introducing above inactivated avec fragment into mcs region of phjl401 . competent cells of et12567 were transformed by recombinant plasmid pid03 and pc05 respectively 以含有avec基因的3 . 1kb基因组dnapsti片段为基础，将1 . 5kb的安普霉素抗性基因片段插入到avec基因中的sphi酶切位点，再将此插入失活的avec基因片段连接到具有接合转移功能(含有orit基因)的链霉菌-大肠杆菌穿梭质粒phjl401的多克隆位点区，由此得到重组质粒pc05 。
Then some necessary elements such as gfp gene , neo gene were inserted between the loxp site and the loxs 11 site and obtained a new gene targeting vector pioxgfp . on the other hand , the chicken v - ifn cdna was cloned into the vector plox between the loxp site and the lox511 site and got another gene targeting vector ploxifn 合成loxp和lox511位点，将其同方向地克隆到载体pegfp - 1的多克隆位点之内，构建成含有lox位点的通用载体? ? plox ，然后将报告基因gfp及neo基因片段克隆到载体plox上的loxp和lox511序列之间，构建成一基因打靶载体? ploxgfp 。
It has been demonstrated directly or indirectly - 7 - that ak auto ab is an important element in the immune network and plays a important role in maintaining physiological functions , clearing aged cells and metabolic products , regulating immune responses and protecting against infection . in some pathological states such as psoriasis and contact dermatitis , a certain serum level of the antibody could inhibit the progression of the diseases , and is beneficial to the recovery from the diseases . after a long time studies on the production and regulation mechanism of physiological and pathological auto antibodies , meanwhile , experiencing an intensive academic debating on whether naas a " horror autotoxicus " or a " gnothi seaution ( know yourself ) " , a common viewpoint has been achieved that naa is of clinical significance in the treatment of immunity diseases for it ' s function in the immune system stability , immunoglobulin y and polyclonal ak auto abs have been used in treating inflammatory dermatitis , and recombinant antibody is under investigating 抗角蛋白自身抗体( akautoab )是naa的重要组成部分，以往实验通过杂交瘤技术、免疫亲和层析技术和噬菌体抗体库技术分别获得单克隆akautoab 、健康人血清多克隆akautoab和基因工程人akautoab ，并对akautoab免疫学特性及在体生理和病理意义进行了广泛的研究，直接或间接地发现akautoab是机体正常免疫调节网络的组成部分，在维护某些生理状态的稳定、清理衰老细胞及代谢产物、调节免疫和抗感染等方面起到重要作用；在某些病理情况下(如银屑病、接触性皮炎等) ，体内akautoab的组分和滴度会发生变化，而正常水平的akautoab则有利于限制病情的发展，促进损伤的修复。
The wells of elisa plate were coated with pab ( 100ng / l ) against h3n2 , then phage was added to the wells . after incubation , the wells were washed vigorously with tbst to remove nonbinding phage . phage bound to the antibody were eluted with 0 . 2mol / l glycine - hcl ( ph2 . 2 ) for 10 min at room temperature and neutrialized with 2mol / l tris - hcl ( ph9 . 1 ) 以抗h3n2流感病毒的多克隆抗体( 100ng l )包被酶标板，加入制备好的肽库，用tbst洗去非特异结合的噬菌体，加0 . 2mol l甘氨酸-盐酸( ph2 . 2 ) ，室温放置10min以洗脱特异结合的噬菌体， 2mol ltris - hcl ( ph9 . 1 )中和后，取2 l噬菌体接种大肠杆菌xl1 - blue菌进行空斑滴定，其余噬菌体扩增后用于下一轮筛选，共重复3轮淘洗。
It acts as a mediator molecule for adhesion between the surface of blastocyst and epithelial cell , and maybe involved in implantation regulation , mmp - 9 is highly expressed and secreted by trophoblast cells , and the antibody of mmp - 9 can inhibit the invasion of the trophoblast this indicates mmp - 9 is very important to the invasive embryo Mmp - 9 (基质金属蛋白酶- 9 )在植入过程中滋养层细胞大量分泌和表达，而多克隆抗mmp - 9抗体可以阻断这些滋养层细胞对基质的浸润，这说明， mmp - 9对于胚胎的侵入至关重要。
In this study , a 1 . 7kb kpni fragment and a lacz gene expression cassette carrying the e . coli lacz gene under the control of sv40 promoter were inserted into the transfer vector pbdtk - uni ( a 277bp acci - accl fragment in the tk gene was deleted ) . the new transfer vector was called puni - lacz . the transfer vector puni - lacz and purified genomic dna of strain bartha - k61 were used to cotransfect vero cells using lipofectin transfection procedure 本研究以呈ge ~ -表型的经典弱毒疫苗bartha - k61株为亲本株，在通用prv转移载体pbdtk - uni的基础上，在其多克隆位点中插入由sv40早期启动子控制下的lacz基因表达盒，同时将下游同源臂增加了一个1 . 7kb的kpni片段，使上下游同源臂的长度都超过了1kb ，构建了一个新的转移载体puni - lacz 。
