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"Latar Belakang: APOBEC3G, apolipoprotein B mRNA-editing enzyme, catalytic polypeplidelike JG, merupakan protein manusia yang dapat mengganggu replikasi HIV dengan memasukkan dirinya ke dalam partikel virus dan merusak susunan materi genetik virus. Beberapa studi terbaru menunjukkan bahwa APOBEC3G manusia mengatur infektivitas HIV-1 dengan mendeaminasi dC menjadi dU pada rantai minus DNA yang baru dibentuk, menyebabkan hipermutasi G menjadi A dari rantai plus DNA viral.Induksi hlpermutasi oleh APOBEC3G dapat menyebabkan pembentukan stop kodon pada ORF protein virus dan memicu degradasi DNA virus oleh glikosilase DNA urnsil yang selanjutuya dapat menghambat replikasi HIV. Protein ini layak untuk diteliti lebih lanjut dalam rangka pengembangan anti retrovirus yang berbasis pacta mekanisme penghambatan replikasi HIV-1 melalui jalur APOBEC3G. Sebagai langkab awal, diperlukan sistem ekspresi gen APOBEC3G yang akan diperoleh melalui sintesis dan kloning gen APOBEC3G ke dalam vektor ekspresi DNA rekombinan.Metode: Untuk mendapatkan mRNA APOBEC3G yang akan digunakan sebagai pola cetak dalam sintesis eDNA APOBEC3G, dilalkukan ekstraksi RNA dari sel CEM-GFP menggunakan Rneasy Mini Kit. Agar DNA APOBEC3G lengkap dapat diperoleh dengan lebih mudah, sintesis DNA serat ganda APOBEC3G menggunakan reaksi RT-PCR dua tahap, dibagi atas tiga daerah pada gen APOBEC3G dengan susunan nukleotida yang bertumpang tindih (overlapping) pada bagian ujung segmen DNA yang akan berfungsi sebagai penyambung fragmen-fragmen tersebut menjadi DNA APOBEC3G utub.Hasil: Hasil eksperimen menunjukloan ketiga fragmen APOBEC3G yang masing-masing berukuran 452 pb, 458 pb,dan 433 pb berhasil dibentuk lewat reaksi PCR dengan menggunakan enzim Pfx dan diklona ke dalam vektor plasmid.Kesimpulan: DNA APOBEC3G yang dibagi menjadi 3 fragmen telah berhasil didapat dan terklona ke dalam pBluescript KS (-). Pekerjaan lanjutan akan dilakukan untuk verifikasi sekuen fragmen-fragmen terklona dan menyambung ketiga fragrnen tersebut menjadi DNA APOBEC3G yang utub yang kemudian akan diklona ke dalam vektor ekspresi.

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Background: APOBEC3G, apolipoprotein B rnRNA-cditing enzyme, catalytic polypeptide-like JG,is a human protein that interferes with the replication of HIV by incorporating itself into virus particles and damaging the genetic material of the virus. Several recent studies revealed that human APOBEC3G regulates HIVI infectivity by dearninating dC to dU in the newly synthesized minus strand DNA, resulting in G to A hypermutation of the viral plus strand DNA.Hypermutation induced by APOBEC3G may result in the introduction of stop codons in viral protein open reading frame and degradation of viral DNA by ura<:il-DNA glyoosylase, therefore blocking HlV replication. This protein is therefore suitable for further investigation for the development of ARV (AntiRetroviral) that is based on mechanism of blocking HIV-1 replication inhibition by APOBEC3G through the pathway. In order to obtain the APOBEC3G protein, an expression system of the APOBEC3G gene is required, which will be obtained by synthesis and cloning of the APOBEC3G gene into an expression vector.Method: To obtain the APOBEC3G mRNA that will be used as template for synthesis of APOBEC3G eDNA by RT-PCR using Omniscript enzyme, we performed RNA extraction from CEM-GFP cell line using the Rneasy Mini !Gt. In order to facilitate the synthesis of a complete APOBEC3G DNA, the APOBEC3G DNA double stranded was divided into three regions with overlapping nucleotide sequences at the DNA ends that function in the joining of the fragments into a full length APOBEC3G DNA.Results: The result of the experiments showed that the three fragments of APOBEC3G gene of 452 bp, 458 bp, and 433 bp, were successfully produced by PCR reaction using Pfx enzyme, and cloned into plasmid vector.Conclusions: APOBEC3G DNA that was divided into three fragments has been obtained and cloned illlo Bluescript KS (-) vector. Further study, will be performed to verifY the cloned fragments, and to fuse the fragments into a complete APOBEC3G DNA that will be cloned into an expression vector."

"Principles of Cloning, Second Edition is the fully revised edition of the authoritative book on the science of cloning. The book presents the basic biological mechanisms of how cloning works and progresses to discuss current and potential applications in basic biology, agriculture, biotechnology, and medicine. Beginning with the history and theory behind cloning, the book goes on to examine methods of micromanipulation, nuclear transfer, genetic modification, and pregnancy and neonatal care of cloned animals. The cloning of various species-including mice, sheep, cattle, and non-mammals-is considered as well. The Editors have been involved in a number of breakthroughs using cloning technique, including the first demonstration that cloning works in differentiated cells done by the Recipient of the 2012 Nobel Prize for Physiology or Medicine - Dr John Gurdon; the cloning of the first mammal from a somatic cell - Drs Keith Campbell and Ian Wilmut; the demonstration that cloning can reset the biological clock - Drs Michael West and Robert Lanza; the demonstration that a terminally differentiated cell can give rise to a whole new individual - Dr Rudolf Jaenisch and the cloning of the first transgenic bovine from a differentiated cell - Dr Jose Cibelli. The majority of the contributing authors are the principal investigators on each of the animal species cloned to date and are expertly qualified to present the state-of-the-art information in their respective areas. First and most comprehensive book on animal cloning, 100% revisedDescribes an in-depth analysis of current limitations of the technology and research areas to exploreOffers cloning applications on basic biology, agriculture, biotechnology, and medicine."

"Describes the miraculous breakthroughs scientists have made in the past decade and dispels the many myths and misunderstandings surrounding stem cell research and therapeutic cloning, offering a window into the astonishing potential of regenerative medicine to extend the human life span and cure disease. Annotation. The extraordinary story of the breakthrough discoveries in cell aging, stem cell research, and therapeutic cloning, and the tremendous promise they hold for dramatically extending human life. Dr. Michael West has been consumed with the mystery of science since he was as an inquisitive child mixing chemicals in his attic-turned-laboratory. Today, he stands in the center of a controversy so great that the list of those lining up against him includes President George W. Bush. Once a devoted creationist eager to dispel theories of human evolution, Dr. West was set on a quest to find a scientific solution to the devastating effects of disease and death after the death of his father. He became immersed in the study of cell aging and the discovery of the cellular "clock" telomerase - the mechanism that controls cell aging. His work led him to found the biotechnology company Geron, a pioneer in the field of stem cell research. His new company, Advanced Cell Technology, is the only organization in the United States pursuing human therapeutic cloning research - research in the field of "regenerative medicine" intended to repair damaged and diseased human organs and tissues. Unlike reproductive cloning, the attempt to clone a human child, therapeutic cloning is a process of growing cells, using a patient's own DNA that is inserted it into an unfertilized egg cell to create embryonic stem cells, cells that hold the promise of repairing the damage of age and disease - in essence, making the cell young again. The potential for therapeutic cloning to treat afflictions caused by the loss of dysfunction of cells - from spinal cord injury and skin burns to kidney failures and cancer - is enormous. Part memoir, part adventure story, The Immortal Cell chronicles the breakthroughs Dr. West and other scientists have made in biotechnology over the past decade - and the astonishing potential they offer us to cure diseases and improve the quality of human life."

"Sebagian besar bakteri asam laktat (BAL) menghasilkan eksopolisakarida (biopolimer fruktan) yang mempunyai banyak manfaat dalam industri makanan, kosmetik, kesehatan dan farmasi. Sintesis biopolimer ini melibatkan peran enzim fruktansukrase atau fruktosiltransferase (ftf). Rekayasa genetika dapat dilakukan untuk mendapatkan biopolimer yang berkriteria unggul, yaitu biopolimer inulin yang mempunyai derajat polimerisasi tinggi. Weissella confusa galur MBFCNC-2(1) telah menjadi sumber gen fruktansukrase yang dikloning lengkap di inang E. coli BL21 StarTM.Tujuan penelitian ini adalah untuk mendapatkan klon versi terpenggal dari gen fruktansukrase karena klon gen lengkap dilaporkan mempunyai masalah dalam ekspresinya. Sebagai template untuk kloning digunakan plasmid dari E. coli BL21 StarTM rekombinan, plasmid rekombinan pO_ftfNS pembawa gen lengkap, dan DNA genomik. PCR dilakukan menggunakan primer FTFdel_Fw dan FTFdel_Rv. Hasil PCR disekuensing dan dianalisis menggunakan BLAST. Sebagai hasil, gen fruktansukrase versi terpenggal didapatkan dari plasmid rekombinan pO_ftfNS pembawa gen ftf lengkap.

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Most of Lactic Acid Bacteria (LAB) produce exopolysaccharide (fructan biopolymer) that has many advantages in food, cosmetic, health, and pharmacy industries. Synthesis of this biopolymer involves the role of fructansucrase enzyme of fructosyltransferase (ftf). Genetic engineering could be done to obtain biopolymer with excellence characteristcs, that is inulin with high degree of polymerization. Weisella confusa strain MBFCNC-2(1) has been used as a source of fructansucrase gene which is full-length clonned at E. coli BL21 StarTM.The aim of this study was to obtain truncated gene of fructansucrase because fulllength clone has problem on its expression. The PCR template used in this study were plasmid of Recombinant E coli BL21 StarTM, recombinant plasmid pO_ftfNS carrying full-length gene, and genomic DNA. PCR was carried out by FTFdel_Fw and FTFdel_Rv primer. The PCR product was sequenced and analyzed by using BLAST. Result revealed that truncated fructansucrase gene was obtained from plasmid recombinant pO_ftfNS carrying full-length gene."

"ABSTRACTActin is a major component of the plant cytoskeleton, so all cells contain this protein. Actin is expressed constitutively and is involved in basic housekeeping functions required for cell maintenance. Because of this, it has been frequently used as an internal control to normalize changes in gene expressions analysis. Actually, the information of nucleotidesequence of actin gene of Jatropha curcas L. population IP-2P from Indonesia is not available yet. The objective of this research was to isolate, clone and characterize cDNA of actin genes of J. curcas IP-2P. Three partial actin gene sequences had been successfully isolated by PCR using total cDNA as template, and actin primer designed from conserved region ofArabidopsis thaliana. Nucleotide sequence analysis showed that the length of JcACT fragment is 610, 534, and 701 bp encoding 203, 177, and 234 amino acids respectively. Local alignment analysis based on mRNA sequences shows that JcACT fragment shares 98% similarity with actin mRNA of Hevea brasiliensis and 99% with actin mRNA of Ricinus communis. Based on deduced amino acid sequence, JcACT is 100% identical to acting from Prunus salicina, Gossypium hirsutum, and Betula luminifera. Even though these clones of cDNA are not completed yet, they can be used as reference in J. curcas L. gene expression analysis."